Metal-based thiophene photodynamic compounds and their use

ABSTRACT

Compositions of the invention include tunable metal-based thiophene photodynamic compounds useful as therapeutic agents and as in vivo diagnostic agents for treating or preventing diseases that involve hyperproliferating cell etiology including cancer and diseases associated with hyperproliferating cells. The compositions are also useful for treating infectious diseases and for pathogen disinfection.

BACKGROUND OF THE INVENTION

1. Field of Invention

This invention relates to photodynamic compounds useful as therapeuticagents and as in vivo diagnostic agents. In particular, the inventionprovides tunable metal-based thiophene photodynamic compounds that canbe activated to cleave DNA upon irradiation with visible light by eithera Type 1 or Type 2 photoprocess.

2. Description of Related Art

Photodynamic therapy (PDT) is currently an active area of research forthe treatment of diseases associated with hyperproliferating cells suchas cancer and non-malignant lesions. PDT has also found use in othercontexts, including but not limited to the treatment of acne, psoriasis,proliferative non-malignant conditions, ulcers and wounds. Thedevelopment ofnew photodynamic compounds (PDCs) (or photosensitizers(PSs)) for photodynamic therapy (PDT) has been increasingly focused onmetallosupramolecular complexes derived from metals such as rutheniumand rhodium. The ongoing investigation of new PSs for PDT stems from thelimitations associated with traditional organic-based porphyrins such asPHOTOFRIN, which must be activated with relatively short wavelengths oflight and do not function in hypoxic environments. Significant advanceshave been made toward overcoming these limitations with the introductionof mixed-metal complexes that possess low-lying ³MMCT (metal-to-metalcharge transfer) excited states. To date, however, there has beenlimited reporting of photodynamic compounds, particularly those with amononuclear or dinuclear design, that are capable of providingphotodynamic therapy for the treatment of diseases associated withhyperproliferating cells such as cancer and non-malignant lesions,and/or capable of treating other conditions including but not limited toinfectious diseases and pathogen infections.

There is a long felt need for new photodynamic compounds (PDCs) that areuseful as photosensitizers for PDT that are both disease-modifying andeffective in treating patients with diseases caused byhyperproliferating cells, for example, cancer. There is also a long feltneed for new PDCs that are useful as in vivo diagnostic agents.Moreover, it is desired to provide novel PDCs having: (1) increasedphoto stability, (2) increased absorption at activation wavelength, (3)red-light, and preferably NIR, absorption, (4) maximal activityregardless of oxygen levels (possibly utilizing a mechanism forswitching between type 1 and type 2 photosensitzation), and (5)intracellular nuclear DNA targeting.

The present invention addresses the need to develop novel PDCs that areuseful as photo sensitizers for PDT that are both disease-modifying andeffective in treating one or more of the conditions discussed above,such as treating patients with diseases caused by hyperproliferatingcells, for example, cancer. The present invention also addresses thelong felt need for new PDCs that are useful as in vivo diagnosticagents.

All references cited herein are incorporated herein by reference intheir entireties.

BRIEF SUMMARY OF THE INVENTION

The present invention is directed toward novel compounds of formula (I),

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein:M is selected from the group consisting of manganese, molybdenum,rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel,platinum, and copper;X is selected from the group consisting of Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄⁻, CF₃SO₃ ⁻, and SO₄ ⁻²;n=0, 1, 2, 3, 4, or 5;y=1, 2, or 3;=0, 1, or 2;Lig at each occurrence is independently selected from the groupconsisting of

R¹ is selected from the group consisting of

u is an integer;R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), and R^(2f) at each occurrenceare each independently selected from the group consisting of hydrogen,C1-6 optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C3-7 optionally substituted cycloalkyl, C1-6 optionallysubstituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶₂, NR⁷ ₂, sulfate, sulfonate, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryl, and optionallysubstituted heterocycle;R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f), R^(3g), R^(3h), R^(3i),R^(3j), R^(3k), R^(3l), and R^(3m) at each occurrence are eachindependently selected from the group consisting of hydrogen, C1-6optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C1-6 optionally substituted halo alkyl, C1-6 optionallysubstituted alkoxy, and CO₂R⁸;R^(4a) and R^(4b) at each occurrence are each independently selectedfrom the group consisting of hydrogen, C1-6 optionally substitutedalkyl, C1-6 optionally substituted branched alkyl, C1-6 optionallysubstituted cycloalkyl, C1-6 optionally substituted halo alkyl, C1-6optionally substituted alkoxy, CO₂R⁵, CONR⁶ ₂, NR⁷ ₂, sulfate,sulfonate, optionally substituted aryl, optionally substituted aryloxy,optionally substituted heteroaryl, and optionally substitutedheterocycle;R^(4a) and R^(4b) at each occurrence on a thiophene ring are takentogether with the atom to which they are bound to form an optionallysubstituted ring having from 6 ring atoms containing 2 oxygen atoms;R⁵ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁶ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁷ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁸ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;

Compounds having the following structures are excluded from certainembodiments of the compounds of formula (I):

The compounds of the present invention include compounds having formula(II),

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein:M is selected from the group consisting ofmanganese, molybdenum,rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel,platinum, and copper;X is selected from the group consisting of Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄⁻, CF₃SO₃ ⁻, and SO₄ ⁻²;n=0, 1, 2, or 3;Lig at each occurrence is independently selected from the groupconsisting of

R¹ is selected from the group consisting of

u is an integer;R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), and R^(2f) at each occurrenceare each independently selected from the group consisting of hydrogen,C1-6 optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C3-7 optionally substituted cycloalkyl, C1-6 optionallysubstituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶₂, NR⁷ ₂, sulfate, sulfonate, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryl, and optionallysubstituted heterocycle;R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3g), R^(3h), R^(3i), R^(3j),R^(3k), R^(3l) and R^(3m) at each occurrence are each independentlyselected from the group consisting of hydrogen, C1-6 optionallysubstituted alkyl, C1-6 optionally substituted branched alkyl, C1-6optionally substituted halo alkyl, C1-6 optionally substituted alkoxy,and CO₂R⁸;R^(4a), R_(4b), and R^(4c) at each occurrence are each independentlyselected from the group consisting of hydrogen, C1-6 optionallysubstituted alkyl, C1-6 optionally substituted branched alkyl, C1-6optionally substituted cycloalkyl, C1-6 optionally substituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶ ₂, NR⁷ ₂,sulfate, sulfonate, optionally substituted aryl, optionally substitutedaryloxy, optionally substituted heteroaryl, and optionally substitutedheterocycle;R^(4a) and R^(4b) at each occurrence on a thiophene ring are takentogether with the atom to which they are bound to form an optionallysubstituted ring having from 6 ring atoms containing 2 oxygen atoms;R⁵ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁶ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁷ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁸ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;

Compounds having the following structures are excluded from certainembodiments of the compounds of formula (II):

The compounds of the present invention includes compounds having formula(III):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof wherein M, Lig, X and the R groups are asdefined above, and g is 0, 1, 2, 3, 4, or 5.

The compounds of the present invention includes compounds having formula(IV):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof wherein M, X and the R groups are asdefined above, and h is 0, 1, 2, 3, 4, or 5.

The present invention is also directed toward novel compounds of formula(V):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein:Lig at each occurrence is independently selected from the groupconsisting of

M at each occurrence is independently selected from the group consistingof manganese, molybdenum, rhenium, iron, ruthenium, osmium, cobalt,rhodium, iridium, nickel, platinum, and copper;X and the R groups are as defined above;t is an integer, and is preferably 1, 2, 3, 4, 5 or 6;q=0, 1, 2, 3, 4 or 5;

The compound having the following structure is excluded from certainembodiments of the compounds of formula (V):

The present invention is also directed toward novel methods of use ofcompounds of the structures

The present invention further relates to compositions comprising aneffective amount of one or more compounds according to the presentinvention and an excipient.

The present invention also relates to a method of using the photodynamiccompounds of the invention as a DNA binding agent.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient as a DNAbinding agent.

The present invention also relates to a method of using the photodynamiccompounds of the invention as a DNA photocleavage agent.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient as a DNAphotocleavage agent.

The present invention also relates to a method of using the photodynamiccompounds of the invention as a DNA condensation agent.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient as DNAcondensation agent.

The present invention also relates to a method of using the photodynamiccompounds of the invention as an agent that produces the DNAlight-switch effect.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient as anagent that produces the DNA light-switch effect.

The present invention also relates to a method of using the photodynamiccompounds of the invention as a photosensitizer with 100% efficiency forsinglet oxygen production.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient as aphotosensitizer with 100% efficiency for singlet oxygen production.

The present invention also relates to a method of using the photodynamiccompounds of the invention as a photosensitizer that can function inboth Type I and Type II photoprocesses.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient as aphotosensitizer that can function in both Type I and Type IIphotoprocesses.

The present invention also relates to a method of using the photodynamiccompounds of the invention to destroy cells, includinghyperproliferating cells.

The present invention also related to a method of using the photodynamiccompounds of the invention to destroy cells, includinghyperproliferating cells, using light as an activator.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient to destroycells, including hyperproliferating cells.

The present invention also relates to a method of using the photodynamiccompounds of the invention to induce apoptosis in cells, includinghyperproliferating cells.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient to induceapoptosis in cells, including hyperproliferating cells.

The present invention also relates to a method of using the photodynamiccompounds of the invention to impart DNA crosslinking in cells,including hyperproliferating cells.

The present invention also relates to a method of using the photodynamiccompounds according to the present invention and an excipient to impartDNA crosslinking in cells, including hyperproliferating cells.

The present invention also relates to a method for treating orpreventing diseases that involve hyperproliferating cells etiology,including, for example, cancer, said method comprising administering toa subject an effective amount of a compound or composition according tothe present invention.

The present invention yet further relates to a method for treating orpreventing diseases that hyperproliferating cells etiology, including,for example, cancer, wherein said method comprises administering to asubject a composition comprising an effective amount of one or morecompounds according to the present invention and an excipient.

The present invention also relates to a method for treating orpreventing diseases that involve hyperproliferating cells in theiretiology, including, for example, cancer, said method comprisingadministering to a subject an effective amount of a compound orcomposition according to the present invention and an intracellularreducing agent such as glutathione.

The present invention also relates to a method for treating orpreventing diseases that involve hyperproliferating cells in theiretiology, including, for example, cancer, said method comprisingadministering to a subject an effective amount of a compound orcomposition according to the present invention and an excipient and anintracellular reducing agent such as glutathione.

The present invention also relates to a method for treating orpreventing diseases that involve hyperproliferating cells in theiretiology, including, for example, cancer, said method comprisingadministering to a subject an effective amount of a compound orcomposition according to the present invention and an intracellularoxidizing agent such as oxygen.

The present invention also relates to a method for treating orpreventing diseases that involve hyperproliferating cells in theiretiology, including, for example, cancer, said method comprisingadministering to a subject an effective amount of a compound orcomposition according to the present invention and an excipient and anintracellular oxidizing agent such as oxygen.

The present invention also relates to a method of using the photodynamiccompounds of the invention as an anti-pathogen medicinal agent anddisinfectant, e.g., to kill microbes in normoxic and hypoxicenvironments.

The present invention also relates to a method of using the photodynamiccompounds of the invention as an in vivo diagnostic agent viaintracellular luminescence.

The present invention further relates to a method of preparing thephotodynamic compounds of the present invention.

These and other objects, features, and advantages will become apparentto those of ordinary skill in the art from a reading of the followingdetailed description and the appended claims. All percentages, ratiosand proportions herein are by weight, unless otherwise specified. Alltemperatures are in degrees Celsius (° C.) unless otherwise specified.All documents cited are in relevant part, incorporated herein byreference; the citation of any document is not to be construed as anadmission that it is prior art with respect to the present invention.

BRIEF DESCRIPTION OF SEVERAL VIEWS OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

The invention will be described in conjunction with the followingdrawings in which like reference numerals designate like elements andwherein:

FIG. 1 shows a set of representative compounds of the disclosure(compounds 1a, 2a, 10a, 10b, 3a, 3b, 14a and 14b).

FIG. 2 is a graph of the UV-Vis absorption spectra of 1a, 10a, and 14ain MeCN. The integrated absorption profiles for 1a, 10a, and 14a in thevisible range (25,000-15,000 cm⁻¹) are 5841, 8190 and 14,100,respectively.

FIG. 3: Light absorption by PDCs 1a, 10a, 14a, and 16a dissolved at 20μM in water.

FIG. 4: is a graph of the ¹O₂ emission sensitized by PDCs 1a, 10a, and14a in MeCN. The ¹O₂ quantum yields for 1a, 10a, and 14a are 0.47, 0.74,and 1.0, respectively.

FIG. 5: (a) Titration of 10a (20 μM) with CT DNA (10 mM Tris·100 mMNaCl, pH 7.5). Inset: binding isotherm at 372 nm. (b) Titration of 10b(50 μM) with CT DNA (10 mM Tris·100 mM NaCl, pH 7.5). Inset: bindingisotherm at 380 nm.

FIG. 6: HL60 cells loaded with 14a viewed by laser scanning confocalmicroscopy. Excitation of the PDC at 458/488 nm produced red emissionthat was collected through a LP510 filter. Images were collected by alaser scanning confocal microscope (Zeiss LSM510 with ZEN operatingsystem) by using 40×/NA1.3, oil or 63×/NA1.4, oil objectives. Excitationwere 458/488 nm of argon/krypton and signals were collected through aLP510 filter. (a) Emission from the PDC 14a, (b) Differentialinterference contract (DIC), and (c) emission/DIC overlay.

FIG. 7: Thermal denaturation of calf-thymus DNA (50 μM, NP) in 5 mM Triswith 50 mM NaCl, pH 7.4) alone and in the presence of 1a, 1b, 10a, and10b (5 μM, [PDC]/[NP]=0.1).

FIG. 8: DNA light-switch effect produced by PDCs: (a) 1a, (b) 10a, and14a.

FIG. 9: Epi-luminescence (EL) and bright field (BF) images ofPDC-treated HL-60 cells: a) untreated control, (b) 1a (100 μM), 5 min.dark (c) 1a (100 μM), 40 hr. dark, and (d) 1a (100 μM), 40 hr.post-irradiation. EL images were collected using a TRITC filter cube(λ_(ex)=540 nm, λ_(em)=605 nm) with no staining. Luminescence observedis from 1a.

FIG. 10: Epi-luminescence (EL) and bright field (BF) images ofPDC-treated HL-60 cells: (a) untreated control, (b) 10a (100 μM), 5 min.dark (c) 10a (100 μM), 40 hr. dark, and (d) 10a (100 μM), 40 hr.post-irradiation. EL images were collected using a TRITC filter cube(λ_(ex)=540 nm, λ_(em)=605 nm) with no staining. Luminescence observedis from 10a.

FIG. 11: Epi-luminescence (EL) and bright field (BF) images ofPDC-treated HL-60 cells: (a) untreated control, (b) 10b (100 μM), 5 min.dark (c) 10b (100 μM), 40 hr. dark, and (d) 10b (100 μM), 40 hr.post-irradiation. EL images were collected using a TRITC filter cube(λ_(ex)=540 nm, λ_(em)=605 nm) with no staining. Luminescence observedis from 10b.

FIG. 12: Epi-luminescence (EL) and bright field (BF) images ofPDC-treated HL-60 cells: (a) untreated control, (b) 14a (100 μM), 5 min.dark (c) 14a (100 μM), 40 hr. dark, and (d) 14a (100 μM), 40 hr.post-irradiation. EL images were collected using a TRITC filter cube(λ_(ex)=540 nm, λ_(em)=605 nm) with no staining. Luminescence observedis from 14a.

FIG. 13: Gel electrophoretic analysis (1% agarose gel containing 0.75 μgmL⁻¹ ethidium bromide, 1×TAE, 8 V cm⁻¹, 30 min.) of DNA photocleavage byPDCs of the disclosure in air-saturated solution with 420-nmirradiation: (a) 1a, (b) 10a, (b) 10b, and (d) 14a. Lanes 1, 2 and 10are control lanes, and lanes 3-9 are reaction lanes: lane 1, 0 μM (−hv);lane 2, 0 μM; lane 3, 0.5 μM; lane 4, 0.75 μM; lane 5, 1.0 μM; lane 6,2.5 μM; lane 7, 5.0 μM; lane 8, 7.5 μM; lane 9, 10 μM; lane 10, 10 μM(−hv).

FIG. 14: Gel electrophoretic analysis (1% agarose gel containing 0.75 μgmL-1 ethidium bromide, 1×TAE, 8 V cm-1, 30 min.) of DNA photocleavage byPDCs in air-saturated solution with visible irradiation: (a) 1a, (b)10a, (b) 10b, and (d) 14a. Lanes 1, 2, 14 and 15 are control lanes, andlanes 2-13 are reaction lanes: lane 1, 0 μM (−hv); lane 2, 0 μM; lane 3,0.75 μM; lane 4, 1.5 μM; lane 5, 3 μM; lane 6, 5 μM; lane 7, 8 μM; lane8, 10 μM; lane 9, 12 μM; lane 10, 16 μM; lane 11, 18 μM; lane 12, 20 μM;lane 13, 25 μM; lane 14, 25 μM (−hv); lane 15, 25 μM (−pUC19).

FIG. 15: Gel electrophoretic analysis (1% agarose gel precast with 0.75μg mL⁻¹ ethidium bromide, 1×TAE, 8 V cm⁻¹, 30 min.) of PDC-mediatedpUC19 photocleavage in air-saturated and deoxygenated solution: 420-nmirradiation of pUC19 (20 μM NP in 10 mM Tris·100 mM NaCl, pH 7.4) for 1hr. Lanes 1 (−PDC, −hv), 2 (300 μM [Ru(bpy)₃]₂₊, −hv), 5 (2 μM 10a,−hv), 8 (4 μM 10b, −hv), and 11 (8 μM 14a, −hv) are dark controls. Lanes3 (300 μM [Ru(bpy)₃]₂₊, +hv), 6 (2 μM 10a, +hv), 9 (4 μM 10b, +hv), and12 (8 μM 14a, +hv) are samples that were irradiated in air; lanes 4, 7,10, and 13 are the corresponding samples irradiated in argon.

FIG. 16: Gel electrophoretic analysis (1% agarose gel precast with 0.75μg mL⁻¹ ethidium bromide, 1×TAE, 8 V cm⁻¹, 30 min.) of PDC-mediatedpUC19 photocleavage in air-saturated and deoxygenated solution: visibleirradiation of pUC19 (20 μM NP in 10 mM Tris·100 mM NaCl, pH 7.4) for 1hr. with cool white fluorescent tubes, 21 W/m². Lanes 1 (−PDC, −hv), 2(500 μM [Ru(bpy)₃]²⁺, −hv), and 5 (2 μM 10a, −hv) are controls. Lanes 3(500 μM [Ru(bpy)₃]²⁺, +hv) and 6 (2 μM 10a, +hv) contain samples thatwere irradiated in air; lanes 4 and 7 are the corresponding samplesirradiated in argon.

FIG. 17: Cytotoxicity (▪) and photocytotoxicity (□) of HL-60 cells (4hr. pre-incubation, 15 min. vis irradiation, 4 J/cm²) with increasingconcentrations of 10a and 10b observed by viability staining with TrypanBlue. (a) 18 hr. and (b) 40 hr. At 100 μM [PDC], the photodynamic actionfor 10a is 47% at 18 hr. and 72% at 40 hr while for 10b it is 11% at 18hr. and 0% (actually 5% increase in viability) at 40 hr. (referencedrelative to 100 μM dark well).

FIG. 18: Cytotoxicity (▪) and photocytotoxicity (□) of HL-60 cells (4hr. pre-incubation, 15 min. vis irradiation 4 J/cm²) with increasingconcentrations of (a) 1a, (b) 10a, and (c) 14a observed by viabilitystaining with Trypan Blue.

FIG. 19: Cytotoxicity and photocytotoxicity of HL-60 cells with varyingconcentrations of 14a observed by nuclear morphology staining withAO-EB. (a) 1 M 14a, −hv; (b) 5 μM 14a, −hv; (c) 10 μM 14a, −hv; (d) 1 μM14a, +hv; (e) 5 μM 14a, +hv; (f) 10 μM 14a, +hv. AO (green fluorescence)stains viable cells; EB (red fluorescence) stains nonviable cells.

FIG. 20: Cytotoxicity (▪) and photocytotoxicity (□) of HL-60 cells (4hr. pre-incubation, 15 min. visible irradiation, 4 J/cm²) withincreasing concentrations of (a) 1a, (b) 10a, and (c) 14a observed byviability staining with Trypan Blue.

FIG. 21: Cytotoxicity (▪) and photocytotoxicity (□) of PDC-treated HL-60cells (4 hr. pre-incubation, 15 min. vis irradiation, 4 J/cm²) withincreasing concentrations of (a) 1a, (b) 10a, and (c) 14a observed byviability staining at 18 hr. post-irradiation with Trypan Blue.

FIG. 22: Cytotoxicity (▪) and photocytotoxicity (□) of PDC-treated HL-60cells (4 hr. pre-incubation, 15 min. vis irradiation, 4 J/cm²) withincreasing concentrations of (a) 1a, (b) 10a, and (c) 14a observed byviability staining at 40 hr. post-irradiation with Trypan Blue.

FIG. 23: Cytotoxicity (▪) and photocytotoxicity (□) of PDC-treated HL-60cells (4 hr. pre-incubation, 15 min. visible irradiation, 4 J/cm²) withincreasing concentrations of (a) 1a, (b) 10a, and (c) 14a observed byviability staining at 18 hr. post-irradiation with Trypan Blue. PDCsprepared by microwave synthesis.

FIG. 24: Cytotoxicity (▪) and photocytotoxicity (□) of PDC-treated HL-60cells (4 hr. pre-incubation, 15 min. visible irradiation, 4 J/cm²) withincreasing concentrations of (a) 1a, (b) 10a, and (c) 14a observed byviability staining at 40 hr. post-irradiation with Trypan Blue. PDCsprepared by microwave synthesis.

FIG. 25: Cytotoxicity (▪) and photocytotoxicity (□) of PDC-treated HL-60cells (4 hr. pre-incubation, 15 min. visible irradiation, 4 J/cm²) withincreasing concentration of 14a at (a) 18 hr. and (b) 40 hr.post-irradiation observed by viability staining with Trypan Blue.

FIG. 26: Formation of apoptotic bodies in HL-60 cells with 100 μM 10a[bottom, left cell in (a)-(c)] observed by nuclear morphology stainingwith AO-EB. (a) Bright-field 130 ms exposure, 100 μM 10a, +hv, 1000×;(b) epi-fluorescence (FITC) 40 ms exposure, 100 μM 10a, +hv, 1000×; (c)epi-fluorescence (TRITC) 40 ms exposure, 100 μM 10a, +hv, 1000×.

FIG. 27: Epi-luminescence (EL) and bright field (BF) apoptotic images ofPDC-treated HL-60 cells at 18 hr. post-irradiation: (a) dark control,14a (5 μM) (b) 14a (5 μM), and (c) 14a (5 μM). EL images were collectedusing a TRITC filter cube (λ_(ex)=540 nm, λ_(em)=605 nm) with EBstaining.

FIG. 28: Bright field (BF) and epi-luminescence (EL) images ofPDC-treated HL-60 cells in the dark [(a)-(b)] and with visibleirradiation [(c)-(f)] observed by nuclear morphology staining with AO-EBat 40 hr. post-irradiation. (a) 5 μM 14a, −hv, BF 1000×; (b) 5 μM 14a,−hv, EL 1000×; (c) 5 μM 14a, +hv, BF 1000×; (d) 5 μM 14a, +hv, EL 1000×;(e) 5 μM 14a, +hv, EL 100×; (f) 5 μM 14a, +hv, EL 100× (zoomed). AO(green fluorescence) stains viable cells; EB (red fluorescence) stainsnonviable cells. Apoptotic bodies are evident in (d) and (f).

FIG. 29: Size distribution of PDC-treated HL-60 cells at 40 hr.post-irradiation for control, 10a, and 10b.

FIG. 30: Size distribution of PDC-treated HL-60 cells at 40 hourspost-irradiation.

FIG. 31: Cyclic voltammogram of 14a.

FIG. 32: Absorption and emission of 14a at 77K.

FIG. 33: DNA photocleavage by increasing concentration of 14a: (a) noGSH, and (b) 4 mM GSH. Lanes 1-10 contain pBR322 plasmid DNA (20 μM-NP)exposed to 14a (0.5-5 μM) and irradiated at 420 nm for 30 min. in 5 mMTris, 50 mM NaCl, pH 7.5.

FIG. 34: The effect of GSH on DNA photodamage increases with increasingn: (a) 1a, n=1, (b) 10a, n=2, and (c) 14a, n=3. Lanes 1-6 and 17-18 arecontrol lanes. Lanes 7-16 contain pBR322 plasmid DNA (20 μM-NP) exposedto PDC (2 μM) and increasing concentration of GSH and irradiated at 420nm for 30 min. in 5 mM Tris, 50 mM NaCl, pH 7.5.

FIG. 35: The effect of AA on DNA photodamage by 14a. Lanes 1-6 and 14-15are control lanes. Lanes 7-13 contain pBR322 plasmid DNA (20 μM-NP)exposed to PDC (2 μM) and increasing concentration of AA and irradiatedat 420 nm for 30 min. in 5 mM Tris, 50 mM NaCl, pH 7.5.

FIG. 36: Formula for determining DNA binding constant for compounds ofthe disclosure.

FIG. 37: Cytotoxicity () and photocytotoxicity (▴) of HL-60 cells (4hr. pre-incubation, 15 min. vis irradiation, 4 J/cm²) with increasingconcentrations of 16a.

FIG. 38: Cytotoxicity () and photocytotoxicity (▴) of HL-60 cells (4hr. pre-incubation, 15 min. vis irradiation, 4 J/cm²) with increasingconcentrations of 16c.

FIG. 39: The efficacy of PDCs 10A, 14A and 16A on cell kill in CT26.WT(top), U87 (middle), and F98 (bottom) cells expressed as the number ofcells killed as a percentage of control (no PDC, no light). The effectsof PDCs without light (dark toxicity, left panels) and with light (PDT,right panels) are shown. Data are shown as means +/− standard errorbars.

FIG. 40: A comparison of the effective doses for PDCs 14A, 14C,PHOTOFRIN, and Aminolevulinic Acid (ALA) used in PDT studies for tumourdestruction in mouse models.

FIG. 41: Cell kill efficacy of PDC 14A versus ALA on CT26.WT (top), U87(middle), and F98 (bottom) cells expressed as the number of cells killedas a percentage of control (no PDC, no light). The effects of PDCwithout light (dark toxicity, left panels) and with light (PDT, rightpanels) are shown.

FIG. 42: The absorption spectra for the photosensitizers 14C (solid) andPPIX (dashed). The wavelength used for PDT experiments is shown by avertical dashed line.

FIG. 43: The absorbance spectra for PDC 14C (square) compared to PPIX(diamond) after 60 minutes of irradiation (525 nm, 78 mW/cm²). ODs wererecorded at the maximal absorbance in the visible region for each PDC,423 nm for 14C, 411 nm for PPIX.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION

Throughout the description, where compositions are described as having,including, or comprising specific components, or where processes aredescribed as having, including, or comprising specific process steps, itis contemplated that compositions of the present teachings also consistessentially of, or consist of, the recited components, and that theprocesses of the present teachings also consist essentially of, orconsist of, the recited processing steps.

In the application, where an element or component is said to be includedin and/or selected from a list of recited elements or components, itshould be understood that the element or component can be any one of therecited elements or components and can be selected from the groupconsisting of two or more of the recited elements or components.

The use of the singular herein includes the plural (and vice versa)unless specifically stated otherwise. In addition, where the use of theterm “about” is before a quantitative value, the present teachings alsoinclude the specific quantitative value itself, unless specificallystated otherwise.

It should be understood that the order of steps or order for performingcertain actions is immaterial so long as the present teachings remainoperable. Moreover, two or more steps or actions can be conductedsimultaneously

As used herein, the term “halogen” shall mean chlorine, bromine,fluorine and iodine.

As used herein, unless otherwise noted, “alkyl” and “aliphatic” whetherused alone or as part of a substituent group refers to straight andbranched carbon chains having 1 to 20 carbon atoms or any number withinthis range, for example 1 to 6 carbon atoms or 1 to 4 carbon atoms.Designated numbers of carbon atoms (e.g. C₁₋₆) shall refer independentlyto the number of carbon atoms in an alkyl moiety or to the alkyl portionof a larger alkyl-containing substituent. Non-limiting examples of alkylgroups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl,iso-butyl, tert-butyl, and the like. Alkyl groups can be optionallysubstituted. Non-limiting examples of substituted alkyl groups includehydroxymethyl, chloromethyl, trifluoromethyl, aminomethyl, 1-chloroethyl, 2-hydroxyethyl, 1,2-difluoroethyl, 3-carboxypropyl, and the like.In substituent groups with multiple alkyl groups such as(C₁₋₆alkyl)₂-amino, the alkyl groups may be the same or different.

As used herein, the terms “alkenyl” and “alkynyl” groups, whether usedalone or as part of a substituent group, refer to straight and branchedcarbon chains having 2 or more carbon atoms, preferably 2 to 20, whereinan alkenyl chain has at least one double bond in the chain and analkynyl chain has at least one triple bond in the chain. Alkenyl andalkynyl groups can be optionally substituted. Nonlimiting examples ofalkenyl groups include ethenyl, 3-propenyl, 1-propenyl (also2-methylethenyl), isopropenyl (also 2-methylethen-2-yl), buten-4-yl, andthe like. Nonlimiting examples of substituted alkenyl groups include2-chloroethenyl (also 2-chlorovinyl), 4-hydroxybuten-1-yl,7-hydroxy-7-methyloct-4-en-2-yl, 7-hydroxy-7-methyloct-3,5-dien-2-yl,and the like. Nonlimiting examples of alkynyl groups include ethynyl,prop-2-ynyl (also propargyl), propyn-1-yl, and 2-methyl-hex-4-yn-1-yl.Nonlimiting examples of substituted alkynyl groups include,5-hydroxy-5-methylhex-3-ynyl, 6-hydroxy-6-methylhept-3-yn-2-yl,5-hydroxy-5-ethylhept-3-ynyl, and the like.

As used herein, “cycloalkyl,” whether used alone or as part of anothergroup, refers to a non-aromatic carbon-containing ring includingcyclized alkyl, alkenyl, and alkynyl groups, e.g., having from 3 to 14ring carbon atoms, preferably from 3 to 7 or 3 to 6 ring carbon atoms,or even 3 to 4 ring carbon atoms, and optionally containing one or more(e.g., 1, 2, or 3) double or triple bond. Cycloalkyl groups can bemonocyclic (e.g., cyclohexyl) or polycyclic (e.g., containing fused,bridged, and/or spiro ring systems), wherein the carbon atoms arelocated inside or outside of the ring system. Any suitable ring positionof the cycloalkyl group can be covalently linked to the defined chemicalstructure. Cycloalkyl rings can be optionally substituted. Nonlimitingexamples of cycloalkyl groups include: cyclopropyl,2-methyl-cyclopropyl, cyclopropenyl, cyclobutyl,2,3-dihydroxycyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl,cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctanyl,decalinyl, 2,5-dimethylcyclopentyl, 3,5-dichlorocyclohexyl,4-hydroxycyclohexyl, 3,3,5-trimethylcyclo hex-1-yl, octahydropentalenyl,octahydro-1H-indenyl, 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl,decahydroazulenyl; bicyclo[6.2.0]decanyl, decahydronaphthalenyl, anddodecahydro-1H-fluorenyl. The term “cycloalkyl” also includes carbocyclic rings which are bicyclic hydrocarbon rings, non-limiting examplesof which include, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl,bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl,bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

“Haloalkyl” is intended to include both branched and straight-chainsaturated aliphatic hydrocarbon groups having the specified number ofcarbon atoms, substituted with 1 or more halogen. Haloalkyl groupsinclude perhaloalkyl groups, wherein all hydrogens of an alkyl grouphave been replaced with halogens (e.g., —CF₃, —CF₂CF₃). Haloalkyl groupscan optionally be substituted with one or more substituents in additionto halogen. Examples of haloalkyl groups include, but are not limitedto, fluoromethyl, dichloroethyl, trifluoromethyl, trichloromethyl,pentafluoroethyl, and pentachloroethyl groups.

The term “alkoxy” refers to the group —O-alkyl, wherein the alkyl groupis as defined above. Alkoxy groups optionally may be substituted. Theterm C₃-C₆ cyclic alkoxy refers to a ring containing 3 to 6 carbon atomsand at least one oxygen atom (e.g., tetrahydrofuran,tetrahydro-2H-pyran). C₃-C₆ cyclic alkoxy groups optionally may besubstituted.

The term “aryl,” wherein used alone or as part of another group, isdefined herein as a an unsaturated, aromatic mono cyclic ring of 6carbon members or to an unsaturated, aromatic polycyclic ring of from 10to 14 carbon members. Aryl rings can be, for example, phenyl or naphthylring each optionally substituted with one or more moieties capable ofreplacing one or more hydrogen atoms. Non-limiting examples of arylgroups include: phenyl, naphthylen-1-yl, naphthylen-2-yl,4-fluorophenyl, 2-hydroxyphenyl, 3-methylphenyl, 2-amino-4-fluorophenyl,2-(N,N-diethylamino)phenyl, 2-cyanophenyl, 2,6-di-tert-butylphenyl,3-methoxyphenyl, 8-hydroxynaphthylen-2-yl 4,5-dimethoxynaphthylen-1-yl,and 6-cyano-naphthylen-1-yl. Aryl groups also include, for example,phenyl or naphthyl rings fused with one or more saturated or partiallysaturated carbon rings (e.g., bicyclo[4.2.0]octa-1,3,5-trienyl,indanyl), which can be substituted at one or more carbon atoms of thearomatic and/or saturated or partially saturated rings.

The term “arylalkyl” or “aralkyl” refers to the group -alkyl-aryl, wherethe alkyl and aryl groups are as defined herein. Aralkyl groups of thepresent invention are optionally substituted. Examples of arylalkylgroups include, for example, benzyl, 1-phenylethyl, 2-phenylethyl,3-phenylpropyl, 2-phenylpropyl, fluorenylmethyl and the like.

The terms “heterocyclic” and/or “heterocycle” and/or “heterocylyl,”whether used alone or as part of another group, are defined herein asone or more ring having from 3 to 20 atoms wherein at least one atom inat least one ring is a heteroatom selected from nitrogen (N), oxygen(O), or sulfur (S), and wherein further the ring that includes theheteroatom is non-aromatic. In heterocycle groups that include 2 or morefused rings, the non-heteroatom bearing ring may be aryl (e.g.,indolinyl, tetrahydroquinolinyl, chromanyl). Exemplary heterocyclegroups have from 3 to 14 ring atoms of which from 1 to 5 are heteroatomsindependently selected from nitrogen (N), oxygen (O), or sulfur (S). Oneor more N or S atoms in a heterocycle group can be oxidized. Heterocyclegroups can be optionally substituted.

Non-limiting examples of heterocyclic units having a single ringinclude: diazirinyl, aziridinyl, urazolyl, azetidinyl, pyrazolidinyl,imidazolidinyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolidinyl,isothiazolyl, isothiazolinyl oxathiazolidinonyl, oxazolidinonyl,hydantoinyl, tetrahydrofuranyl, pyrrolidinyl, morpholinyl, piperazinyl,piperidinyl, dihydropyranyl, tetrahydropyranyl, piperidin-2-onyl(valerolactam), 2,3,4,5-tetrahydro-1H-azepinyl, 2,3-dihydro-1H-indo le,and 1,2,3,4-tetrahydro-quinoline. Non-limiting examples of heterocyclicunits having 2 or more rings include: hexahydro-1H-pyrrolizinyl,3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl,3a,4,5,6,7,7a-hexahydro-1H-indolyl, 1,2,3,4-tetrahydroquinolinyl,chromanyl, isochromanyl, indolinyl, isoindolinyl, anddecahydro-1H-cycloocta[b]pyrrolyl.

The term “heteroaryl,” whether used alone or as part of another group,is defined herein as one or more rings having from 5 to 20 atoms whereinat least one atom in at least one ring is a heteroatom chosen fromnitrogen (N), oxygen (O), or sulfur (S), and wherein further at leastone of the rings that includes a heteroatom is aromatic. In heteroarylgroups that include 2 or more fused rings, the non-heteroatom bearingring may be a carbocycle (e.g., 6,7-Dihydro-5H-cyclopentapyrimidine) oraryl (e.g., benzofuranyl, benzothiophenyl, indolyl). Exemplaryheteroaryl groups have from 5 to 14 ring atoms and contain from 1 to 5ring heteroatoms independently selected from nitrogen (N), oxygen (O),or sulfur (S). One or more N or S atoms in a heteroaryl group can beoxidized. Heteroaryl groups can be substituted. Non-limiting examples ofheteroaryl rings containing a single ring include: 1,2,3,4-tetrazolyl,[1,2,3]triazolyl, [1,2,4]triazolyl, triazinyl, thiazolyl, 1H-imidazolyl,oxazolyl, furanyl, thiopheneyl, pyrimidinyl, 2-phenylpyrimidinyl,pyridinyl, 3-methylpyridinyl, and 4-dimethylaminopyridinyl. Non-limitingexamples of heteroaryl rings containing 2 or more fused rings include:benzofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl,benztriazolyl, cinnolinyl, naphthyridinyl, phenanthridinyl, 7H-purinyl,9H-purinyl, 6-amino-9H-purinyl, 5H-pyrrolo[3,2-d]pyrimidinyl,7H-pyrrolo[2,3-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl,2-phenylbenzo[d]thiazolyl, 1H-indolyl, 4,5,6,7-tetrahydro-1-H-indolyl,quinoxalinyl, 5-methylquinoxalinyl, quinazolinyl, quinolinyl,8-hydroxy-quinolinyl, and isoquinolinyl.

One non-limiting example of a heteroaryl group as described above isC₁-C₅ heteroaryl, which has 1 to 5 carbon ring atoms and at least oneadditional ring atom that is a heteroatom (preferably 1 to 4 additionalring atoms that are heteroatoms) independently selected from nitrogen(N), oxygen (O), or sulfur (S). Examples of C₁-C₅ heteroaryl include,but are not limited to, triazinyl, thiazol-2-yl, thiazol-4-yl,imidazol-1-yl, 1H-imidazol-2-yl, 1H-imidazol-4-yl, isoxazolin-5-yl,furan-2-yl, furan-3-yl, thiophen-2-yl, thiophen-4-yl, pyrimidin-2-yl,pyrimidin-4-yl, pyrimidin-5-yl, pyridin-2-yl, pyridin-3-yl, andpyridin-4-yl.

Unless otherwise noted, when two substituents are taken together to forma ring having a specified number of ring atoms (e.g., R² and R³ takentogether with the nitrogen (N) to which they are attached to form a ringhaving from 3 to 7 ring members), the ring can have carbon atoms andoptionally one or more (e.g., 1 to 3) additional heteroatomsindependently selected from nitrogen (N), oxygen (O), or sulfur (S). Thering can be saturated or partially saturated and can be optionallysubstituted.

For the purposed of the present invention fused ring units, as well asspirocyclic rings, bicyclic rings and the like, which comprise a singleheteroatom will be considered to belong to the cyclic familycorresponding to the heteroatom containing ring. For example,1,2,3,4-tetrahydroquinoline having the formula:

is, for the purposes of the present invention, considered a heterocyclicunit. 6,7-Dihydro-5H-cyclopentapyrimidine having the formula:

is, for the purposes of the present invention, considered a heteroarylunit. When a fused ring unit contains heteroatoms in both a saturatedand an aryl ring, the aryl ring will predominate and determine the typeof category to which the ring is assigned. For example,1,2,3,4-tetrahydro-[1,8]naphthyridine having the formula:

is, for the purposes of the present invention, considered a heteroarylunit.

Whenever a term or either of their prefix roots appear in a name of asubstituent the name is to be interpreted as including those limitationsprovided herein. For example, whenever the term “alkyl” or “aryl” oreither of their prefix roots appear in a name of a substituent (e.g.,arylalkyl, alkylamino) the name is to be interpreted as including thoselimitations given above for “alkyl” and “aryl.”

The term “substituted” is used throughout the specification. The term“substituted” is defined herein as a moiety, whether acyclic or cyclic,which has one or more hydrogen atoms replaced by a substituent orseveral (e.g., 1 to 10) substituents as defined herein below. Thesubstituents are capable of replacing one or two hydrogen atoms of asingle moiety at a time. In addition, these substituents can replace twohydrogen atoms on two adjacent carbons to form said substituent, newmoiety or unit. For example, a substituted unit that requires a singlehydrogen atom replacement includes halogen, hydroxyl, and the like. Atwo hydrogen atom replacement includes carbonyl, oximino, and the like.A two hydrogen atom replacement from adjacent carbon atoms includesepoxy, and the like. The term “substituted” is used throughout thepresent specification to indicate that a moiety can have one or more ofthe hydrogen atoms replaced by a substituent. When a moiety is describedas “substituted” any number of the hydrogen atoms may be replaced. Forexample, difluoromethyl is a substituted C₁ alkyl; trifluoromethyl is asubstituted C₁ alkyl; 4-hydroxyphenyl is a substituted aromatic ring;(N,N-dimethyl-5-amino)octanyl is a substituted C₈ alkyl;3-guanidinopropyl is a substituted C₃ alkyl; and 2-carboxypyridinyl is asubstituted heteroaryl.

The variable groups defined herein, e.g., alkyl, alkenyl, alkynyl,cycloalkyl, alkoxy, aryloxy, aryl, heterocycle and heteroaryl groupsdefined herein, whether used alone or as part of another group, can beoptionally substituted. Optionally substituted groups will be soindicated.

The following are non-limiting examples of substituents which cansubstitute for hydrogen atoms on a moiety: halogen (chlorine (Cl),bromine (Br), fluorine (F) and iodine (I)), —CN, —NO₂, oxo (═O), —OR⁹,—SR⁹, —N(R⁹)₂, —NR⁹C(O)R⁹, —SO₂R⁹, —SO₂OR⁹, —SO₂N(R⁹)₂, —C(O)R⁹,—C(O)OR⁹, —C(O)N(R⁹)₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₂₋₈alkenyl, C₂₋₈ alkynyl, C₃₋₁₄ cycloalkyl, aryl, heterocycle, orheteroaryl, wherein each of the alkyl, haloalkyl, alkenyl, alkynyl,alkoxy, cycloalkyl, aryl, heterocycle, and heteroaryl groups isoptionally substituted with 1-10 (e.g., 1-6 or 1-4) groups selectedindependently from halogen, —CN, —NO₂, oxo, and R^(x); wherein R^(x), ateach occurrence, independently is hydrogen, —OR¹⁰, —SR¹⁰, —C(O)R¹⁰,—C(O)OR¹⁰, —C(O)N(R¹⁰)₂, —SO₂R¹⁰, —S(O)₂OR¹⁰, —N(R¹⁰)₂, —NR¹⁰C(O)R¹⁰,C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, cycloalkyl(e.g., C₃₋₆ cycloalkyl), aryl, heterocycle, or heteroaryl, or two R^(x)units taken together with the atom(s) to which they are bound form anoptionally substituted carbocycle or heterocycle wherein said carbocycleor heterocycle has 3 to 7 ring atoms; wherein R¹⁰, at each occurrence,independently is hydrogen, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, cycloalkyl (e.g., C₃₋₆ cycloalkyl), aryl, heterocycle, orheteroaryl, or two R¹⁰ units taken together with the atom(s) to whichthey are bound form an optionally substituted carbocycle or heterocyclewherein said carbocycle or heterocycle preferably has 3 to 7 ring atoms.

In certain embodiments, the substituents are selected from

-   -   i) —OR¹¹; for example, —OH, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃;    -   ii) —C(O)R¹¹; for example, —COCH₃, —COCH₂CH₃, —COCH₂CH₂CH₃;    -   iii) —C(O)OR¹¹; for example, —CO₂CH₃, —CO₂CH₂CH₃, —CO₂CH₂CH₂CH₃;    -   iv) —C(O)N(R¹¹)₂; for example, —CONH₂, —CONHCH₃, —CON(CH₃)₂;    -   v) —N(R¹¹)₂; for example, —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃);    -   vi) halogen: —F, —Cl, —Br, and —I;    -   vii) —CH_(e)X_(g); wherein X is halogen, m is from 0 to 2,        e+g=3; for example, —CH₂F, —CHF₂, —CF₃, —CCl₃, or —CBr₃;    -   viii) —SO₂R¹¹; for example, —SO₂H; —SO₂CH₃; —SO₂C₆H₅;    -   ix) C₁-C₆ linear, branched, or cyclic alkyl;    -   x) Cyano    -   xi) Nitro;    -   xii) N(R¹¹)C(O)R¹¹;    -   xiii) Oxo (═O);    -   xiv) Heterocycle; and    -   xv) Heteroaryl.        wherein each R¹¹ is independently hydrogen, optionally        substituted C₁-C₆ linear or branched alkyl (e.g., optionally        substituted C₁-C₄ linear or branched alkyl), or optionally        substituted C₃-C₆ cycloalkyl (e.g optionally substituted C₃-C₄        cycloalkyl); or two R¹¹ units can be taken together to form a        ring comprising 3-7 ring atoms. In certain aspects, each R¹¹ is        independently hydrogen, C₁-C₆ linear or branched alkyl        optionally substituted with halogen or C₃-C₆ cycloalkyl or C₃-C₆        cycloalkyl.

At various places in the present specification, substituents ofcompounds are disclosed in groups or in ranges. It is specificallyintended that the description include each and every individualsubcombination of the members of such groups and ranges. For example,the term “C₁₋₆ alkyl” is specifically intended to individually discloseC₁, C₂, C₃, C₄, C₅, C₆, C₁-C₆, C₁-C₅, C₁-C₄, C₁-C₃, C₁-C₂, C₂-C₆, C₂-C₅,C₂-C₄, C₂-C₃, C₃-C₆, C₃-C₅, C₃-C₄, C₄-C₆, C₄-C₅, and C₅-C₆, alkyl.

For the purposes of the present invention the terms “compound,”“analog,” and “composition of matter” stand equally well for thephotodynamic compounds described herein, including all enantiomericforms, diastereomeric forms, salts, and the like, and the terms“compound,” “analog,” and “composition of matter” are usedinterchangeably throughout the present specification.

For the purposes of the present invention the term “bpy” will standequally well for 2,2′-bipyridine and [2,2′]bipyridine.

For the purposes of the present invention the term “phen” will standequally well for [1,10]phenanthroline and 1,10-phenanthroline.

For the purposes of the present invention the term “dmb” will standequally well for 4,4′-dimethyl-2,2′-bipyridine.

Compounds described herein can contain an asymmetric atom (also referredas a chiral center), and some of the compounds can contain one or moreasymmetric atoms or centers, which can thus give rise to optical isomers(enantiomers) and diastereomers. The present teachings and compoundsdisclosed herein include such enantiomers and diastereomers, as well asthe racemic and resolved, enantiomerically pure R and S stereoisomers,as well as other mixtures of the R and S stereoisomers andpharmaceutically acceptable salts thereof. Optical isomers can beobtained in pure form by standard procedures known to those skilled inthe art, which include, but are not limited to, diastereomeric saltformation, kinetic resolution, and asymmetric synthesis. The presentteachings also encompass cis and trans isomers of compounds containingalkenyl moieties (e.g., alkenes and imines). It is also understood thatthe present teachings encompass all possible regioisomers, and mixturesthereof, which can be obtained in pure form by standard separationprocedures known to those skilled in the art, and include, but are notlimited to, column chromatography, thin-layer chromatography, andhigh-performance liquid chromatography.

Pharmaceutically acceptable salts of compounds of the present teachings,which can have an acidic moiety, can be formed using organic andinorganic bases. Both mono and polyanionic salts are contemplated,depending on the number of acidic hydrogens available for deprotonation.Suitable salts formed with bases include metal salts, such as alkalimetal or alkaline earth metal salts, for example sodium, potassium, ormagnesium salts; ammonia salts and organic amine salts, such as thoseformed with morpholine, thiomorpholine, piperidine, pyrrolidine, amono-, di- or tri-lower alkylamine (e.g., ethyl-tert-butyl-, diethyl-,diisopropyl-, triethyl-, tributyl- or dimethylpropylamine), or a mono-,di-, or trihydroxy lower alkylamine (e.g., mono-, di- ortriethanolamine). Specific non-limiting examples of inorganic basesinclude NaHCO₃, Na₂CO₃, KHCO₃, K₂CO₃, Cs₂CO₃, LiOH, NaOH, KOH, NaH₂PO₄,Na₂HPO₄, and Na₃PO₄. Internal salts also can be formed. Similarly, whena compound disclosed herein contains a basic moiety, salts can be formedusing organic and inorganic acids. For example, salts can be formed fromthe following acids: acetic, propionic, lactic, benzenesulfonic,benzoic, camphorsulfonic, citric, tartaric, succinic, dichloroacetic,ethenesulfonic, formic, fumaric, gluconic, glutamic, hippuric,hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, malonic,mandelic, methanesulfonic, mucic, napthalenesulfonic, nitric, oxalic,pamoic, pantothenic, phosphoric, phthalic, propionic, succinic,sulfuric, tartaric, toluenesulfonic, and camphorsulfonic as well asother known pharmaceutically acceptable acids.

When any variable occurs more than one time in any constituent or in anyformula, its definition in each occurrence is independent of itsdefinition at every other occurrence (e.g., in N(R⁶)₂, each R⁶ may bethe same or different than the other). Combinations of substituentsand/or variables are permissible only if such combinations result instable compounds.

The terms “treat” and “treating” and “treatment” as used herein, referto partially or completely alleviating, inhibiting, ameliorating and/orrelieving a condition from which a patient is suspected to suffer.

As used herein, “therapeutically effective” and “effective dose” referto a substance or an amount that elicits a desirable biological activityor effect.

As used herein, the term “photodynamic therapy” shall mean a treatmentfor destroying cells and tissue through use of a drug that can beactivated by light of a certain wavelength and dose.

As used herein, the term “photodynamic compound” shall mean a compoundthat provides photodynamic therapy.

Except when noted, the terms “subject” or “patient” are usedinterchangeably and refer to mammals such as human patients andnon-human primates, as well as experimental animals such as rabbits,rats, and mice, and other animals. Accordingly, the term “subject” or“patient” as used herein means any mammalian patient or subject to whichthe compounds of the invention can be administered. In an exemplaryembodiment of the present invention, to identify subject patients fortreatment according to the methods of the invention, accepted screeningmethods are employed to determine risk factors associated with atargeted or suspected disease or condition or to determine the status ofan existing disease or condition in a subject. These screening methodsinclude, for example, conventional work-ups to determine risk factorsthat may be associated with the targeted or suspected disease orcondition. These and other routine methods allow the clinician to selectpatients in need of therapy using the methods and compounds of thepresent invention.

The Photodynamic Compounds

The photodynamic compounds of the present invention are tunablemetal-based thiophenes, and include all enantiomeric and diastereomericforms and pharmaceutically accepted salts thereof having the formula(I),

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein:M is selected from the group consisting ofmanganese, molybdenum,rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel,platinum, and copper;X is selected from the group consisting of Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄⁻, CF₃SO₃ ⁻, and SO₄ ⁻²;n=0, 1, 2, 3, 4, or 5;y=1, 2, or 3;z=0, 1, or 2;Lig at each occurrence is independently selected from the groupconsisting of

R¹ is selected from the group consisting of

u is an integer, and in certain embodiments, is 1-1000 or 1-500 or 1-100or 1-10, or is at least 2 or at least 3 or at least 4 or at least 5 orat least 10, or is any integer from 1-5 or 1-10 or 1-20, or any value upto the point where processability becomes problematic due to aggregationand/or insolubility;R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), and R^(2f) at each occurrenceare each independently selected from the group consisting of hydrogen,C1-6 optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C3-7 optionally substituted cycloalkyl, C1-6 optionallysubstituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶₂, NR⁷ ₂, sulfate, sulfonate, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryl, and optionallysubstituted heterocycle;R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f), R^(3g), R^(3h), R^(3i),R^(3j), R^(3k), R^(3l) and R^(3m) at each occurrence are eachindependently selected from the group consisting of hydrogen, C1-6optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C1-6 optionally substituted halo alkyl, C1-6 optionallysubstituted alkoxy, and CO₂R⁸;R^(4a), R^(4b), and R^(4c) at each occurrence are each independentlyselected from the group consisting of hydrogen, C1-6 optionallysubstituted alkyl, C1-6 optionally substituted branched alkyl, C1-6optionally substituted cyclo alkyl, C1-6 optionally substituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶ ₂, NR⁷ ₂,sulfate, sulfonate, optionally substituted aryl, optionally substitutedaryloxy, optionally substituted heteroaryl, and optionally substitutedheterocycle;R^(4a) and R^(4b) at each occurrence on a thiophene ring are takentogether with the atom to which they are bound to form an optionallysubstituted ring having from 6 ring atoms containing 2 oxygen atoms;R⁵ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁶ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁷ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁸ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl.

Compounds of the structures

are excluded from the novel compounds of formula (I).

The compounds of the present invention includes compounds having formula(II),

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein:M is selected from the group consisting ofmanganese, molybdenum,rhenium, iron, ruthenium, osmium, cobalt, rhodium, iridium, nickel,platinum, and copper;X is selected from the group consisting of Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄⁻, CF₃SO₃ ⁻, and SO₄ ⁻²;n=0, 1, 2 or 3;Lig at each occurrence is independently selected from the groupconsisting of

R¹ is selected from the group consisting of

u is an integer, and in certain embodiments, is 1-1000 or 1-500 or 1-100or 1-10, or is at least 2 or at least 3 or at least 4 or at least 5 orat least 10, or is any integer from 1-5 or 1-10 or 1-20, or any value upto the point where processability becomes problematic due to aggregationand/or insolubility;R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), and R^(2f) at each occurrenceare each independently selected from the group consisting of hydrogen,C1-6 optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C3-7 optionally substituted cycloalkyl, C1-6 optionallysubstituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶₂, NR⁷ ₂, sulfate, sulfonate, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryl, and optionallysubstituted heterocycle;R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f), R^(3g), R^(3h), R^(3i),R^(3j), R^(3k), R^(3l) and R^(3m) at each occurrence are eachindependently selected from the group consisting of hydrogen, C1-6optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C1-6 optionally substituted halo alkyl, C1-6 optionallysubstituted alkoxy, and CO₂R⁸;R^(4a), R^(4b), and R^(4c) at each occurrence are each independentlyselected from the group consisting of hydrogen, C1-6 optionallysubstituted alkyl, C1-6 optionally substituted branched alkyl, C1-6optionally substituted cycloalkyl, C1-6 optionally substituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶ ₂, NR⁷ ₂,sulfate, sulfonate, optionally substituted aryl, optionally substitutedaryloxy, optionally substituted heteroaryl, and optionally substitutedheterocycle;R^(4a) and R^(4b) at each occurrence on a thiophene ring are takentogether with the atom to which they are bound to form an optionallysubstituted ring having from 6 ring atoms containing 2 oxygen atoms;R⁵ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁶ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁷ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;R⁸ at each occurrence is independently selected from the groupconsisting of hydrogen and optionally substituted alkyl;

Compounds of the structures

are excluded from the novel compounds of formula (II).

The compounds of the present invention includes compounds having formula(III),

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof wherein M, Lig, X and the R groups are asdefined above, and g is 0, 1, 2, 3, 4, or 5.

The present is also directed toward novel compounds of formula (IV):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof wherein M, X and the R groups are asdefined above, and h is 0, 1, 2, 3, 4, or 5.

The present invention is also directed toward novel compounds of formula(V):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein:Lig at each occurrence is independently selected from the groupconsisting of

M at each occurrence is independently selected from the group consistingof manganese, molybdenum, rhenium, iron, ruthenium, osmium, cobalt,rhodium, iridium, nickel, platinum, and copper;X and the R groups are as defined above;t is an integer, and is preferably 1, 2, 3, 4, 5 or 6;q=0, 1, 2, 3, 4 or 5.

The compound having the following structure is excluded from certainembodiments of the compounds of formula (V):

The present invention is also directed toward novel methods of use ofcompounds of the structure

In certain embodiments, M is manganese molybdenum, rhenium, iron,ruthenium, osmium, cobalt, rhodium, iridium, nickel, platinum, orcopper.

In certain embodiments, X is Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄ ⁻, CF₃SO₃ ⁻,or SO₄ ⁻².

In certain embodiments, n is 0 or 1 or 2 or 3 or 4 or 5.

In certain embodiments, y is 1 or 2 or 3.

In certain embodiments, z is 0 or 1 or 2.

In certain embodiments, g is 0 or 1 or 2 or 3 or 4 or 5.

In certain embodiments, h is 0 or 1 or 2 or 3 or 4 or 5.

In certain embodiments, t is an integer.

In certain embodiments, t is 1 or 2 or 3 or 4 or 5 or 6.

In certain embodiments, q is 0 or 1 or 2 or 3 or 4 or 5.

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, Lig is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, u is an integer, and in certain of theseembodiments, is 1-1000 or 1-500 or 1-100 or 1-10, or is at least 2 or atleast 3 or at least 4 or at least 5 or at least 10, or is any integerfrom 1-5 or 1-10 or 1-20, or any value up to the point whereprocessability becomes problematic due to aggregation and/orinsolubility.

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R^(2a) is hydrogen.

In certain embodiments, R^(2a) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(2a) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(2a) is C3-7 optionally substitutedcycloalkyl.

In certain embodiments, R^(2a) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(2a) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(2a) is CO₂R⁵.

In certain embodiments, R^(2b) is CONR⁶ ₂.

In certain embodiments, R^(2a) is NR⁷ ₂.

In certain embodiments, R^(2a) is sulfate.

In certain embodiments, R^(2a) is sulfonate.

In certain embodiments, R^(2a) is optionally substituted aryl.

In certain embodiments, R^(2a) is optionally substituted aryloxy.

In certain embodiments, R^(2a) is optionally substituted heteroaryl.

In certain embodiments, R^(2a) is optionally substituted heterocycle.

In certain embodiments, R^(2b) is hydrogen.

In certain embodiments, R^(2b) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(2b) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(2b) is C3-7 optionally substitutedcycloalkyl.

In certain embodiments, R^(2b) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(2b) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(2b) is CO₂R⁵.

In certain embodiments, R^(2b) is CONR⁶ ₂.

In certain embodiments, R^(2b) is NR⁷ ₂.

In certain embodiments, R^(2b) is sulfate.

In certain embodiments, R^(2b) is sulfonate.

In certain embodiments, R^(2b) is optionally substituted aryl.

In certain embodiments, R^(2b) is optionally substituted aryloxy.

In certain embodiments, R^(2b) is optionally substituted heteroaryl.

In certain embodiments, R^(2a) is optionally substituted heterocycle.

In certain embodiments, R^(2c) is hydrogen.

In certain embodiments, R^(2c) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(2c) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(2c) is C3-7 optionally substitutedcycloalkyl.

In certain embodiments, R^(2c) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(2c) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(2c) is CO₂R⁵.

In certain embodiments, R^(2c) is CONR⁶ ₂.

In certain embodiments, R^(2c) is NR⁷ ₂.

In certain embodiments, R^(2c) is sulfate.

In certain embodiments, R^(2c) is sulfonate.

In certain embodiments, R^(2c) is optionally substituted aryl.

In certain embodiments, R^(2c) is optionally substituted aryloxy.

In certain embodiments, R^(2c) is optionally substituted heteroaryl.

In certain embodiments, R^(2c) is optionally substituted heterocycle.

In certain embodiments, R^(2d) is hydrogen.

In certain embodiments, R^(2d) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(2d) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(2d) is C3-7 optionally substitutedcycloalkyl.

In certain embodiments, R^(2d) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(2d) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(2d) is CO₂R⁵.

In certain embodiments, R^(2d) is CONR⁶ ₂.

In certain embodiments, R^(2d) is NR⁷ ₂.

In certain embodiments, R^(2d) is sulfate.

In certain embodiments, R^(2d) is sulfonate.

In certain embodiments, R^(2d) is optionally substituted aryl.

In certain embodiments, R^(2d) is optionally substituted aryloxy.

In certain embodiments, R^(2d) is optionally substituted heteroaryl.

In certain embodiments, R^(2d) is optionally substituted heterocycle.

In certain embodiments, R^(2e) is hydrogen.

In certain embodiments, R^(2e) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(2e) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(2e) is C3-7 optionally substitutedcycloalkyl.

In certain embodiments, R^(2e) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(2e) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(2e) is CO₂R⁵.

In certain embodiments, R^(2e) is CONR⁶ ₂.

In certain embodiments, R^(2e) is NR⁷ ₂.

In certain embodiments, R^(2e) is sulfate.

In certain embodiments, R^(2e) is sulfonate.

In certain embodiments, R^(2e) is optionally substituted aryl.

In certain embodiments, R^(2e) is optionally substituted aryloxy.

In certain embodiments, R^(2e) is optionally substituted heteroaryl.

In certain embodiments, R^(2e) is optionally substituted heterocycle.

In certain embodiments, R^(2f) is hydrogen.

In certain embodiments, R^(2f) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(2f) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(2f) is C3-7 optionally substitutedcycloalkyl.

In certain embodiments, R^(2f) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(2f) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(2f) is CO₂R⁵.

In certain embodiments, R^(2f) is CONR⁶ ₂.

In certain embodiments, R^(2f) is NR⁷ ₂.

In certain embodiments, R^(2f) is sulfate.

In certain embodiments, R^(2f) is sulfonate.

In certain embodiments, R^(2f) is optionally substituted aryl.

In certain embodiments, R^(2f) is optionally substituted aryloxy.

In certain embodiments, R^(2f) is optionally substituted heteroaryl.

In certain embodiments, R^(2f) is optionally substituted heterocycle.

In certain embodiments, R^(3a) is hydrogen.

In certain embodiments, R^(3a) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3a) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3a) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3a) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3a) is CO₂R⁸.

In certain embodiments, R^(3b) is hydrogen.

In certain embodiments, R^(3b) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3b) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3b) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3b) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3b) is CO₂R⁸.

In certain embodiments, R^(3c) is hydrogen.

In certain embodiments, R^(3c) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3c) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3c) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3c) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3c) is CO₂R⁸.

In certain embodiments, R^(3d) is hydrogen.

In certain embodiments, R^(3d) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3d) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3d) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3d) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3d) is CO₂R⁸.

In certain embodiments, R^(3e) is hydrogen.

In certain embodiments, R^(3e) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3e) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3e) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3e) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3e) is CO₂R⁸.

In certain embodiments, R^(3f) is hydrogen.

In certain embodiments, R^(3f) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3f) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3f) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3f) C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3f) is CO₂R⁸.

In certain embodiments, R^(3g) is hydrogen.

In certain embodiments, R^(3g) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3g) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3g) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3g) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3g) is CO₂R⁸.

In certain embodiments, R^(3h) is hydrogen.

In certain embodiments, R^(3h) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3h) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3h) is C1-6 optionally substituted haloalkyl

In certain embodiments, R^(3h) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3h) is CO₂R⁸.

In certain embodiments, R^(3i) is hydrogen.

In certain embodiments, R^(3i) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3i) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3i) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3i) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3i) is CO₂R⁸.

In certain embodiments, R^(3i) is hydrogen.

In certain embodiments, R^(3j) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3j) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3j) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3j) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3j) is CO₂R⁸.

In certain embodiments, R^(3k) is hydrogen.

In certain embodiments, R^(3k) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3k) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3k) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3k) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3k) is CO₂R⁸.

In certain embodiments, R^(3l) is hydrogen.

In certain embodiments, R^(3l) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3l) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3l) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3l) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3l) is CO₂R⁸.

In certain embodiments, R^(3m) is hydrogen.

In certain embodiments, R^(3m) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(3m) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(3m) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(3m) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(3m) is CO₂R⁸.

In certain embodiments, R^(4a) is hydrogen.

In certain embodiments, R^(4a) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(4a) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(4a) is C1-6 optionally substitutedcycloalkyl.

In certain embodiments, R^(4a) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(4a) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(4a) is CO₂R⁵.

In certain embodiments, R^(4a) is CONR⁶ ₂

In certain embodiments, R^(4a) is NR⁷ ₂.

In certain embodiments, R^(4a) is sulfate.

In certain embodiments, R^(4a) is sulfonate.

In certain embodiments, R^(4a) is optionally substituted aryl.

In certain embodiments, R^(4a) is optionally substituted aryloxy.

In certain embodiments, R^(4a) is optionally substituted heteroaryl.

In certain embodiments, R^(4a) is optionally substituted heterocycle.

In certain embodiments, R^(4b) is hydrogen.

In certain embodiments, R^(4b) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(4b) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(4b) is C1-6 optionally substitutedcycloalkyl.

In certain embodiments, R^(4b) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(4b) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(4b) is CO₂R⁵.

In certain embodiments, R^(4b) is CONR⁶ ₂

In certain embodiments, R^(4b) is NR⁷ ₂.

In certain embodiments, R^(4b) is sulfate.

In certain embodiments, R^(4b) is sulfonate.

In certain embodiments, R^(4b) is optionally substituted aryl.

In certain embodiments, R^(4b) is optionally substituted aryloxy.

In certain embodiments, R^(4b) is optionally substituted heteroaryl.

In certain embodiments, R^(4b) is optionally substituted heterocycle.

In certain embodiments, R^(4c) is hydrogen.

In certain embodiments, R^(4c) is C1-6 optionally substituted alkyl.

In certain embodiments, R^(4c) is C1-6 optionally substituted branchedalkyl.

In certain embodiments, R^(4c) is C1-6 optionally substitutedcycloalkyl.

In certain embodiments, R^(4c) is C1-6 optionally substituted haloalkyl.

In certain embodiments, R^(4c) is C1-6 optionally substituted alkoxy.

In certain embodiments, R^(4c) is CO₂R⁵.

In certain embodiments, R^(4c) is CONR⁶ ₂

In certain embodiments, R^(4c) is NR⁷ ₂.

In certain embodiments, R^(4c) is sulfate.

In certain embodiments, R^(4c) is sulfonate.

In certain embodiments, R^(4c) is optionally substituted aryl.

In certain embodiments, R^(4c) is optionally substituted aryloxy.

In certain embodiments, R^(4c) is optionally substituted heteroaryl.

In certain embodiments, R^(4c) is optionally substituted heterocycle.

In certain embodiments, R^(4a) and R^(4b) are taken together with theatom to which they are bound to form an optionally substituted ringhaving from 6 ring atoms containing 2 oxygen atoms.

In certain embodiments, R⁵ is hydrogen.

In certain embodiments, R⁵ is optionally substituted alkyl.

In certain embodiments, R⁶ is hydrogen.

In certain embodiments, R⁶ is optionally substituted alkyl.

In certain embodiments, R⁷ is hydrogen.

In certain embodiments, R⁷ is optionally substituted alkyl.

In certain embodiments, R⁸ is hydrogen.

In certain embodiments, R⁸ is optionally substituted alkyl.

In certain embodiments, q is 0 or 1 or 2 or 3 or 4 or 5.

In certain embodiments, t is 1 or 2 or 3 or 4 or 5 or 6.

Exemplary embodiments include compounds having the formula (VI) or apharmaceutically acceptable salt form thereof:

wherein non-limiting examples of M, Lig, and R¹ are defined herein belowin Table 1.

TABLE 1 Entry M Lig R¹ X n  1 Ru

Cl 2  2 Ru

Cl 2  3 Ru

Cl 2  4 Ru

Cl 2  5 Ru

Cl 2  6 Ru

Cl 2  7 Ru

Cl 2  8 Ru

Cl 2  9 Ru

Cl 2  10 Ru

Cl 2  11 Ru

Cl 2  12 Ru

Cl 2  13 Ru

Cl 2  14 Ru

Cl 2  15 Ru

Cl 2  16 Ru

Cl 2  17 Ru

Cl 2  18 Ru

Cl 2  19 Ru

Cl 2  20 Ru

Cl 2  21 Ru

Cl 2  22 Ru

Cl 2  23 Ru

Cl 2  24 Ru

Cl 2  25 Ru

Cl 2  26 Ru

Cl 2  27 Ru

Cl 2  28 Ru

Cl 2  29 Ru

Cl 2  30 Ru

Cl 2  31 Ru

Cl 2  32 Ru

Cl 2  33 Ru

Cl 2  34 Ru

Cl 2  35 Ru

Cl 2  36 Ru

Cl 2  37 Ru

Cl 2  38 Ru

Cl 2  39 Ru

Cl 2  40 Ru

Cl 2  41 Ru

Cl 2  42 Ru

Cl 2  43 Ru

Cl 2  44 Ru

Cl 2  45 Ru

Cl 2  46 Ru

Cl 2  47 Ru

Cl 2  48 Ru

Cl 2  49 Ru

Cl 2  50 Ru

Cl 2  51 Ru

Cl 2  52 Ru

Cl 2  53 Ru

Cl 2  54 Ru

Cl 2  55 Ru

Cl 2  56 Ru

Cl 2  57 Ru

Cl 2  58 Ru

Cl 2  59 Ru

Cl 2  60 Ru

Cl 2  61 Ru

Cl 2  62 Ru

Cl 2  63 Ru

Cl 2  64 Ru

Cl 2  65 Ru

Cl 2  66 Ru

Cl 2  67 Ru

Cl 2  68 Ru

Cl 2  69 Ru

Cl 2  70 Ru

Cl 2  71 Ru

Cl 2  72 Ru

Cl 2  73 Ru

Cl 2  74 Ru

Cl 2  75 Ru

Cl 2  76 Ru

Cl 2  77 Ru

Cl 2  78 Ru

Cl 2  79 Ru

Cl 2  80 Ru

Cl 2  81 Ru

Cl 2  82 Ru

Cl 2  83 Ru

Cl 2  84 Ru

Cl 2  85 Ru

Cl 2  86 Ru

Cl 2  87 Ru

Cl 2  88 Ru

Cl 2  89 Ru

Cl 2  90 Ru

Cl 2  91 Ru

Cl 2  92 Ru

Cl 2  93 Ru

Cl 2  94 Ru

Cl 2  95 Ru

Cl 2  96 Ru

Cl 2  97 Ru

Cl 2  98 Ru

Cl 2  99 Ru

Cl 2 100 Ru

Cl 2 101 Ru

Cl 2 102 Ru

Cl 2 103 Ru

Cl 2 104 Ru

Cl 2 105 Ru

Cl 2 106 Ru

Cl 2 107 Ru

Cl 2 108 Ru

Cl 2 109 Ru

Cl 2 110 Ru

Cl 2 111 Ru

Cl 2 112 Ru

Cl 2 113 Ru

Cl 2 114 Ru

Cl 2 115 Ru

Cl 2 116 Ru

Cl 2 117 Ru

Cl 2 118 Ru

Cl 2 119 Ru

Cl 2 120 Ru

Cl 2 121 Ru

Cl 2 122 Ru

Cl 2 123 Ru

Cl 2 124 Ru

Cl 2 125 Ru

Cl 2 126 Ru

Cl 2 127 Ru

Cl 2 128 Ru

Cl 2 129 Ru

Cl 2 130 Ru

Cl 2 131 Ru

Cl 2 132 Ru

Cl 2 133 Ru

Cl 2 134 Ru

Cl 2 135 Ru

Cl 2 136 Ru

Cl 2 137 Ru

Cl 2 138 Ru

Cl 2 139 Ru

Cl 2 140 Ru

Cl 2 141 Ru

Cl 2 142 Ru

Cl 2 143 Ru

Cl 2 144 Ru

Cl 2 145 Ru

Cl 2 146 Ru

Cl 2 147 Ru

Cl 2 148 Ru

Cl 2 149 Ru

Cl 2 150 Ru

Cl 2 151 Ru

Cl 2 152 Ru

Cl 2 153 Ru

Cl 2 154 Ru

Cl 2 155 Ru

Cl 2 156 Ru

Cl 2 157 Ru

Cl 2 158 Ru

Cl 2 159 Ru

Cl 2 160 Ru

Cl 2 161 Ru

Cl 2 162 Ru

Cl 2 163 Ru

Cl 2 164 Ru

Cl 2 165 Ru

Cl 2 166 Ru

Cl 2 167 Ru

Cl 2 168 Ru

Cl 2 169 Ru

Cl 2 170 Ru

Cl 2 171 Ru

Cl 2 172 Ru

Cl 2 173 Ru

Cl 2 174 Ru

Cl 2 175 Ru

Cl 2 176 Ru

Cl 2 177 Ru

Cl 2 178 Ru

Cl 2 179 Ru

Cl 2 180 Ru

Cl 2 181 Ru

Cl 2 182 Ru

Cl 2 183 Ru

Cl 2 184 Ru

Cl 2 185 Ru

Cl 2 186 Ru

Cl 2 187 Ru

Cl 2 188 Ru

Cl 2 189 Ru

Cl 2 190 Ru

Cl 2 191 Ru

Cl 2 192 Ru

Cl 2 193 Ru

Cl 2 194 Ru

Cl 2 195 Ru

Cl 2 196 Ru

Cl 2 197 Ru

Cl 2 198 Ru

Cl 2 199 Ru

Cl 2 200 Ru

Cl 2 201 Ru

Cl 2 202 Ru

Cl 2 203 Ru

Cl 2 204 Ru

Cl 2 205 Ru

PF₆ 2 206 Ru

PF₆ 2 207 Ru

PF₆ 2 208 Ru

PF₆ 2 209 Ru

PF₆ 2 210 Ru

PF₆ 2 211 Ru

PF₆ 2 212 Ru

PF₆ 2 213 Ru

PF₆ 2 214 Ru

PF₆ 2 215 Ru

PF₆ 2 216 Ru

PF₆ 2 217 Ru

PF₆ 2 218 Ru

PF₆ 2 219 Ru

PF₆ 2 220 Ru

PF₆ 2 221 Ru

PF₆ 2 222 Ru

PF₆ 2 223 Ru

PF₆ 2 224 Ru

PF₆ 2 225 Ru

PF₆ 2 226 Ru

PF₆ 2 227 Ru

PF₆ 2 228 Ru

PF₆ 2 229 Ru

PF₆ 2 230 Ru

PF₆ 2 231 Ru

PF₆ 2 232 Ru

PF₆ 2 233 Ru

PF₆ 2 234 Ru

PF₆ 2 235 Ru

PF₆ 2 236 Ru

PF₆ 2 237 Ru

PF₆ 2 238 Ru

PF₆ 2 239 Ru

PF₆ 2 240 Ru

PF₆ 2 241 Ru

PF₆ 2 242 Ru

PF₆ 2 243 Ru

PF₆ 2 244 Ru

PF₆ 2 245 Ru

PF₆ 2 246 Ru

PF₆ 2 247 Ru

PF₆ 2 248 Ru

PF₆ 2 249 Ru

PF₆ 2 250 Ru

PF₆ 2 251 Ru

PF₆ 2 252 Ru

PF₆ 2 253 Ru

PF₆ 2 254 Ru

PF₆ 2 255 Ru

PF₆ 2 256 Ru

PF₆ 2 257 Ru

PF₆ 2 258 Ru

PF₆ 2 259 Ru

PF₆ 2 260 Ru

PF₆ 2 261 Ru

PF₆ 2 262 Ru

PF₆ 2 263 Ru

PF₆ 2 264 Ru

PF₆ 2 265 Ru

PF₆ 2 266 Ru

PF₆ 2 267 Ru

PF₆ 2 268 Ru

PF₆ 2 269 Ru

PF₆ 2 270 Ru

PF₆ 2 271 Ru

PF₆ 2 272 Ru

PF₆ 2 273 Ru

PF₆ 2 274 Ru

PF₆ 2 275 Ru

PF₆ 2 276 Ru

PF₆ 2 277 Ru

PF₆ 2 278 Ru

PF₆ 2 279 Ru

PF₆ 2 280 Ru

PF₆ 2 281 Ru

PF₆ 2 282 Ru

PF₆ 2 283 Ru

PF₆ 2 284 Ru

PF₆ 2 285 Ru

PF₆ 2 286 Ru

PF₆ 2 287 Ru

PF₆ 2 288 Ru

PF₆ 2 289 Ru

PF₆ 2 290 Ru

PF₆ 2 291 Ru

PF₆ 2 292 Ru

PF₆ 2 293 Ru

PF₆ 2 294 Ru

PF₆ 2 295 Ru

PF₆ 2 296 Ru

PF₆ 2 297 Ru

PF₆ 2 298 Ru

PF₆ 2 299 Ru

PF₆ 2 300 Ru

PF₆ 2 301 Ru

PF₆ 2 302 Ru

PF₆ 2 303 Ru

PF₆ 2 304 Ru

PF₆ 2 305 Ru

PF₆ 2 306 Ru

PF₆ 2 307 Ru

PF₆ 2 308 Ru

PF₆ 2 309 Ru

PF₆ 2 310 Ru

PF₆ 2 311 Ru

PF₆ 2 312 Ru

PF₆ 2 313 Ru

PF₆ 2 314 Ru

PF₆ 2 315 Ru

PF₆ 2 316 Ru

PF₆ 2 317 Ru

PF₆ 2 318 Ru

PF₆ 2 319 Ru

PF₆ 2 320 Ru

PF₆ 2 321 Ru

PF₆ 2 322 Ru

PF₆ 2 323 Ru

PF₆ 2 324 Ru

PF₆ 2 325 Ru

PF₆ 2 326 Ru

PF₆ 2 327 Ru

PF₆ 2 328 Ru

PF₆ 2 329 Ru

PF₆ 2 330 Ru

PF₆ 2 331 Ru

PF₆ 2 332 Ru

PF₆ 2 333 Ru

PF₆ 2 334 Ru

PF₆ 2 335 Ru

PF₆ 2 336 Ru

PF₆ 2 337 Ru

PF₆ 2 338 Ru

PF₆ 2 339 Ru

PF₆ 2 340 Ru

PF₆ 2 341 Ru

PF₆ 2 342 Ru

PF₆ 2 343 Ru

PF₆ 2 344 Ru

PF₆ 2 345 Ru

PF₆ 2 346 Ru

PF₆ 2 347 Ru

PF₆ 2 348 Ru

PF₆ 2 349 Ru

PF₆ 2 350 Ru

PF₆ 2 351 Ru

PF₆ 2 352 Ru

PF₆ 2 353 Ru

PF₆ 2 354 Ru

PF₆ 2 355 Ru

PF₆ 2 356 Ru

PF₆ 2 357 Ru

PF₆ 2 358 Ru

PF₆ 2 359 Ru

PF₆ 2 360 Ru

PF₆ 2 361 Ru

PF₆ 2 362 Ru

PF₆ 2 363 Ru

PF₆ 2 364 Ru

PF₆ 2 365 Ru

PF₆ 2 366 Ru

PF₆ 2 367 Ru

PF₆ 2 368 Ru

PF₆ 2 369 Ru

PF₆ 2 370 Ru

PF₆ 2 371 Ru

PF₆ 2 372 Ru

PF₆ 2 373 Ru

PF₆ 2 374 Ru

PF₆ 2 375 Ru

PF₆ 2 376 Ru

PF₆ 2 377 Ru

PF₆ 2 378 Ru

PF₆ 2 379 Ru

PF₆ 2 380 Ru

PF₆ 2 381 Ru

PF₆ 2 382 Ru

PF₆ 2 383 Ru

PF₆ 2 384 Ru

PF₆ 2 385 Ru

PF₆ 2 386 Ru

PF₆ 2 387 Ru

PF₆ 2 388 Ru

PF₆ 2 389 Ru

PF₆ 2 390 Ru

PF₆ 2 391 Ru

PF₆ 2 392 Ru

PF₆ 2 393 Ru

PF₆ 2 394 Ru

PF₆ 2 395 Ru

PF₆ 2 396 Ru

PF₆ 2 397 Ru

PF₆ 2 398 Ru

PF₆ 2 399 Ru

PF₆ 2 400 Ru

PF₆ 2 401 Ru

PF₆ 2 402 Ru

PF₆ 2 403 Ru

PF₆ 2 404 Ru

PF₆ 2 405 Ru

PF₆ 2 406 Ru

PF₆ 2 407 Ru

PF₆ 2 408 Ru

PF₆ 2

Exemplary embodiments include compounds having the formula (VII) or apharmaceutically acceptable salt form thereof:

wherein non-limiting examples of M, Lig, and R1 are defined herein belowin Table 2.

TABLE 2 Entry M Lig R¹ X G  1 Ru

Cl 2  2 Ru

Cl 2  3 Ru

Cl 2  4 Ru

Cl 2  5 Ru

Cl 2  6 Ru

Cl 2  7 Ru

Cl 2  8 Ru

Cl 2  9 Ru

Cl 2  10 Ru

Cl 2  11 Ru

Cl 2  12 Ru

Cl 2  13 Ru

Cl 2  14 Ru

Cl 2  15 Ru

Cl 2  16 Ru

Cl 2  17 Ru

Cl 2  18 Ru

Cl 2  19 Ru

Cl 2  20 Ru

Cl 2  21 Ru

Cl 2  22 Ru

Cl 2  23 Ru

Cl 2  24 Ru

Cl 2  25 Ru

Cl 2  26 Ru

Cl 2  27 Ru

Cl 2  28 Ru

Cl 2  29 Ru

Cl 2  30 Ru

Cl 2  31 Ru

Cl 2  32 Ru

Cl 2  33 Ru

Cl 2  34 Ru

Cl 2  35 Ru

Cl 2  36 Ru

Cl 2  37 Ru

Cl 2  38 Ru

Cl 2  39 Ru

Cl 2  40 Ru

Cl 2  41 Ru

Cl 2  42 Ru

Cl 2  43 Ru

Cl 2  44 Ru

Cl 2  45 Ru

Cl 2  46 Ru

Cl 2  47 Ru

Cl 2  48 Ru

Cl 2  49 Ru

Cl 2  50 Ru

Cl 2  51 Ru

Cl 2  52 Ru

Cl 2  53 Ru

Cl 2  54 Ru

Cl 2  55 Ru

Cl 2  56 Ru

Cl 2  57 Ru

Cl 2  58 Ru

Cl 2  59 Ru

Cl 2  60 Ru

Cl 2  61 Ru

Cl 2  62 Ru

Cl 2  63 Ru

Cl 2  64 Ru

Cl 2  65 Ru

Cl 2  66 Ru

Cl 2  67 Ru

Cl 2  68 Ru

Cl 2  69 Ru

Cl 2  70 Ru

Cl 2  71 Ru

Cl 2  72 Ru

Cl 2  73 Ru

Cl 2  74 Ru

Cl 2  75 Ru

Cl 2  76 Ru

Cl 2  77 Ru

Cl 2  78 Ru

Cl 2  79 Ru

Cl 2  80 Ru

Cl 2  81 Ru

Cl 2  82 Ru

Cl 2  83 Ru

Cl 2  84 Ru

Cl 2  85 Ru

Cl 2  86 Ru

Cl 2  87 Ru

Cl 2  88 Ru

Cl 2  89 Ru

Cl 2  90 Ru

Cl 2  91 Ru

Cl 2  92 Ru

Cl 2  93 Ru

Cl 2  94 Ru

Cl 2  95 Ru

Cl 2  96 Ru

Cl 2  97 Ru

Cl 2  98 Ru

Cl 2  99 Ru

Cl 2 100 Ru

Cl 2 101 Ru

Cl 2 102 Ru

Cl 2 103 Ru

Cl 2 104 Ru

Cl 2 105 Ru

Cl 2 106 Ru

Cl 2 107 Ru

Cl 2 108 Ru

Cl 2 109 Ru

Cl 2 110 Ru

Cl 2 111 Ru

Cl 2 112 Ru

Cl 2 113 Ru

Cl 2 114 Ru

Cl 2 115 Ru

Cl 2 116 Ru

Cl 2 117 Ru

Cl 2 118 Ru

Cl 2 119 Ru

Cl 2 120 Ru

Cl 2 121 Ru

Cl 2 122 Ru

Cl 2 123 Ru

Cl 2 124 Ru

Cl 2 125 Ru

Cl 2 126 Ru

Cl 2 127 Ru

Cl 2 128 Ru

Cl 2 129 Ru

Cl 2 130 Ru

Cl 2 131 Ru

Cl 2 132 Ru

Cl 2 133 Ru

Cl 2 134 Ru

Cl 2 135 Ru

Cl 2 136 Ru

Cl 2 137 Ru

Cl 2 138 Ru

Cl 2 139 Ru

Cl 2 140 Ru

Cl 2 141 Ru

Cl 2 142 Ru

Cl 2 143 Ru

Cl 2 144 Ru

Cl 2 145 Ru

Cl 2 146 Ru

Cl 2 147 Ru

Cl 2 148 Ru

Cl 2 149 Ru

Cl 2 150 Ru

Cl 2 151 Ru

Cl 2 152 Ru

Cl 2 153 Ru

Cl 2 154 Ru

Cl 2 155 Ru

Cl 2 156 Ru

Cl 2 157 Ru

Cl 2 158 Ru

Cl 2 159 Ru

Cl 2 160 Ru

Cl 2 161 Ru

Cl 2 162 Ru

Cl 2 163 Ru

Cl 2 164 Ru

Cl 2 165 Ru

Cl 2 166 Ru

Cl 2 167 Ru

Cl 2 168 Ru

Cl 2 169 Ru

Cl 2 170 Ru

Cl 2 171 Ru

Cl 2 172 Ru

Cl 2 173 Ru

Cl 2 174 Ru

Cl 2 175 Ru

Cl 2 176 Ru

Cl 2 177 Ru

Cl 2 178 Ru

Cl 2 179 Ru

Cl 2 180 Ru

Cl 2 181 Ru

Cl 2 182 Ru

Cl 2 183 Ru

Cl 2 184 Ru

Cl 2 185 Ru

Cl 2 186 Ru

Cl 2 187 Ru

Cl 2 188 Ru

Cl 2 189 Ru

Cl 2 190 Ru

Cl 2 191 Ru

Cl 2 192 Ru

Cl 2 193 Ru

Cl 2 194 Ru

Cl 2 195 Ru

Cl 2 196 Ru

Cl 2 197 Ru

Cl 2 198 Ru

Cl 2 199 Ru

Cl 2 200 Ru

Cl 2 201 Ru

Cl 2 202 Ru

Cl 2 203 Ru

Cl 2 204 Ru

Cl 2 205 Ru

PF₆ 2 206 Ru

PF₆ 2 207 Ru

PF₆ 2 208 Ru

PF₆ 2 209 Ru

PF₆ 2 210

PF₆ 2 211 Ru

PF₆ 2 212 Ru

PF₆ 2 213 Ru

PF₆ 2 214 Ru

PF₆ 2 215 Ru

PF₆ 2 216 Ru

PF₆ 2 217 Ru

PF₆ 2 218 Ru

PF₆ 2 219 Ru

PF₆ 2 220 Ru

PF₆ 2 221 Ru

PF₆ 2 222 Ru

PF₆ 2 223 Ru

PF₆ 2 224 Ru

PF₆ 2 225 Ru

PF₆ 2 226 Ru

PF₆ 2 227 Ru

PF₆ 2 228 Ru

PF₆ 2 229 Ru

PF₆ 2 230 Ru

PF₆ 2 231 Ru

PF₆ 2 232 Ru

PF₆ 2 233 Ru

PF₆ 2 234 Ru

PF₆ 2 235 Ru

PF₆ 2 236 Ru

PF₆ 2 237 Ru

PF₆ 2 238 Ru

PF₆ 2 239 Ru

PF₆ 2 240 Ru

PF₆ 2 241 Ru

PF₆ 2 242 Ru

PF₆ 2 243 Ru

PF₆ 2 244 Ru

PF₆ 2 245 Ru

PF₆ 2 246 Ru

PF₆ 2 247 Ru

PF₆ 2 248 Ru

PF₆ 2 249 Ru

PF₆ 2 250 Ru

PF₆ 2 251 Ru

PF₆ 2 252 Ru

PF₆ 2 253 Ru

PF₆ 2 254 Ru

PF₆ 2 255 Ru

PF₆ 2 256 Ru

PF₆ 2 257 Ru

PF₆ 2 258 Ru

PF₆ 2 259 Ru

PF₆ 2 260 Ru

PF₆ 2 261 Ru

PF₆ 2 262 Ru

PF₆ 2 263 Ru

PF₆ 2 264 Ru

PF₆ 2 265 Ru

PF₆ 2 266 Ru

PF₆ 2 267 Ru

PF₆ 2 268 Ru

PF₆ 2 269 Ru

PF₆ 2 270 Ru

PF₆ 2 271 Ru

PF₆ 2 272 Ru

PF₆ 2 273 Ru

PF₆ 2 274 Ru

PF₆ 2 275 Ru

PF₆ 2 276 Ru

PF₆ 2 277 Ru

PF₆ 2 278 Ru

PF₆ 2 279 Ru

PF₆ 2 280 Ru

PF₆ 2 281 Ru

PF₆ 2 282 Ru

PF₆ 2 283 Ru

PF₆ 2 284 Ru

PF₆ 2 285 Ru

PF₆ 2 286 Ru

PF₆ 2 287 Ru

PF₆ 2 288 Ru

PF₆ 2 289 Ru

PF₆ 2 290 Ru

PF₆ 2 291 Ru

PF₆ 2 292 Ru

PF₆ 2 293 Ru

PF₆ 2 294 Ru

PF₆ 2 295 Ru

PF₆ 2 296 Ru

PF₆ 2 297 Ru

PF₆ 2 298 Ru

PF₆ 2 299 Ru

PF₆ 2 300 Ru

PF₆ 2 301 Ru

PF₆ 2 302 Ru

PF₆ 2 303 Ru

PF₆ 2 304 Ru

PF₆ 2 305 Ru

PF₆ 2 306 Ru

PF₆ 2 307 Ru

PF₆ 2 308 Ru

PF₆ 2 309 Ru

PF₆ 2 310 Ru

PF₆ 2 311 Ru

PF₆ 2 312 Ru

PF₆ 2 313 Ru

PF₆ 2 314 Ru

PF₆ 2 315 Ru

PF₆ 2 316 Ru

PF₆ 2 317 Ru

PF₆ 2 318 Ru

PF₆ 2 319 Ru

PF₆ 2 320 Ru

PF₆ 2 321 Ru

PF₆ 2 322 Ru

PF₆ 2 323 Ru

PF₆ 2 324 Ru

PF₆ 2 325 Ru

PF₆ 2 326 Ru

PF₆ 2 327 Ru

PF₆ 2 328 Ru

PF₆ 2 329 Ru

PF₆ 2 330 Ru

PF₆ 2 331 Ru

PF₆ 2 332 Ru

PF₆ 2 333 Ru

PF₆ 2 334 Ru

PF₆ 2 335 Ru

PF₆ 2 336 Ru

PF₆ 2 337 Ru

PF₆ 2 338 Ru

PF₆ 2 339 Ru

PF₆ 2 340 Ru

PF₆ 2 341 Ru

PF₆ 2 342 Ru

PF₆ 2 343 Ru

PF₆ 2 344 Ru

PF₆ 2 345 Ru

PF₆ 2 346 Ru

PF₆ 2 347 Ru

PF₆ 2 348 Ru

PF₆ 2 349 Ru

PF₆ 2 350 Ru

PF₆ 2 351 Ru

PF₆ 2 352 Ru

PF₆ 2 353 Ru

PF₆ 2 354 Ru

PF₆ 2 355 Ru

PF₆ 2 356 Ru

PF₆ 2 357 Ru

PF₆ 2 358 Ru

PF₆ 2 359 Ru

PF₆ 2 360 Ru

PF₆ 2 361 Ru

PF₆ 2 362 Ru

PF₆ 2 363 Ru

PF₆ 2 364 Ru

PF₆ 2 365 Ru

PF₆ 2 366 Ru

PF₆ 2 367 Ru

PF₆ 2 368 Ru

PF₆ 2 369 Ru

PF₆ 2 370 Ru

PF₆ 2 371 Ru

PF₆ 2 372 Ru

PF₆ 2 373 Ru

PF₆ 2 374 Ru

PF₆ 2 375 Ru

PF₆ 2 376 Ru

PF₆ 2 377 Ru

PF₆ 2 378 Ru

PF₆ 2 379 Ru

PF₆ 2 380 Ru

PF₆ 2 381 Ru

PF₆ 2 382 Ru

PF₆ 2 383 Ru

PF₆ 2 384 Ru

PF₆ 2 385 Ru

PF₆ 2 386 Ru

PF₆ 2 387 Ru

PF₆ 2 388 Ru

PF₆ 2 389 Ru

PF₆ 2 390 Ru

PF₆ 2 391 Ru

PF₆ 2 392 Ru

PF₆ 2 393 Ru

PF₆ 2 394 Ru

PF₆ 2 395 Ru

PF₆ 2 396 Ru

PF₆ 2 397 Ru

PF₆ 2 398 Ru

PF₆ 2 399 Ru

PF₆ 2 400 Ru

PF₆ 2 401 Ru

PF₆ 2 402 Ru

PF₆ 2 403 Ru

PF₆ 2 404 Ru

PF₆ 2 405 Ru

PF₆ 2 406 Ru

PF₆ 2 407 Ru

PF₆ 2 408 Ru

PF₆ 2

Exemplary embodiments include compounds having the formula (VIII) or apharmaceutically acceptable salt form thereof:

wherein non-limiting examples of M, R¹, X, and h are defined hereinbelow in Table 3.

TABLE 3 Entry M R¹ X h  1 Ru

Cl 2  2 Ru

Cl 2  3 Ru

Cl 2  4 Ru

Cl 2  5 Ru

Cl 2  6 Ru

Cl 2  7 Ru

Cl 2  8 Ru

Cl 2  9 Ru

Cl 2  10 Ru

Cl 2  11 Ru

Cl 2  12 Ru

Cl 2  13 Ru

Cl 2  14 Ru

Cl 2  15 Ru

Cl 2  16 Ru

Cl 2  17 Ru

Cl 2  18 Ru

Cl 2  19 Ru

Cl 2  20 Ru

Cl 2  21 Ru

Cl 2  22 Ru

Cl 2  23 Ru

Cl 2  24 Ru

Cl 2  25 Ru

Cl 2  26 Ru

Cl 2  27 Ru

Cl 2  28 Ru

Cl 2  29 Ru

Cl 2  30 Ru

Cl 2  31 Ru

Cl 2  32 Ru

Cl 2  33 Ru

Cl 2  34 Ru

Cl 2  35 Ru

Cl 2  36 Ru

Cl 2  37 Ru

Cl 2  38 Ru

Cl 2  39 Ru

Cl 2  40 Ru

Cl 2  41 Ru

Cl 2  42 Ru

Cl 2  43 Ru

Cl 2  44 Ru

Cl 2  45 Ru

Cl 2  46 Ru

Cl 2  47 Ru

Cl 2  48 Ru

Cl 2  49 Ru

Cl 2  50 Ru

Cl 2  51 Ru

Cl 2  52 Ru

Cl 2  53 Ru

Cl 2  54 Ru

Cl 2  55 Ru

Cl 2  56 Ru

Cl 2  57 Ru

Cl 2  58 Ru

Cl 2  59 Ru

Cl 2  60 Ru

Cl 2  61 Ru

Cl 2  62 Ru

Cl 2  63 Ru

Cl 2  64 Ru

Cl 2  65 Ru

Cl 2  66 Ru

Cl 2  67 Ru

Cl 2  68 Ru

Cl 2  69 Ru

Cl 2  70 Ru

Cl 2  71 Ru

Cl 2  72 Ru

Cl 2  73 Ru

Cl 2  74 Ru

Cl 2  75 Ru

Cl 2  76 Ru

Cl 2  77 Ru

Cl 2  78 Ru

Cl 2  79 Ru

Cl 2  80 Ru

Cl 2  81 Ru

Cl 2  82 Ru

Cl 2  83 Ru

Cl 2  84 Ru

Cl 2  85 Ru

Cl 2  86 Ru

Cl 2  87 Ru

Cl 2  88 Ru

Cl 2  89 Ru

Cl 2  90 Ru

Cl 2  91 Ru

Cl 2  92 Ru

Cl 2  93 Ru

Cl 2  94 Ru

Cl 2  95 Ru

Cl 2  96 Ru

Cl 2  97 Ru

Cl 2  98 Ru

Cl 2  99 Ru

Cl 2 100 Ru

Cl 2 101 Ru

Cl 2 102 Ru

Cl 2 103 Ru

Cl 2 104 Ru

Cl 2 105 Ru

Cl 2 106 Ru

Cl 2 107 Ru

Cl 2 108 Ru

Cl 2 109 Ru

Cl 2 110 Ru

Cl 2 111 Ru

Cl 2 112 Ru

Cl 2 113 Ru

Cl 2 114 Ru

Cl 2 115 Ru

Cl 2 116 Ru

Cl 2 117 Ru

Cl 2 118 Ru

Cl 2 119 Ru

Cl 2 120 Ru

Cl 2 121 Ru

Cl 2 122 Ru

Cl 2 123 Ru

Cl 2 124 Ru

Cl 2 125 Ru

Cl 2 126 Ru

Cl 2 127 Ru

Cl 2 128 Ru

Cl 2 129 Ru

Cl 2 130 Ru

Cl 2 131 Ru

Cl 2 132 Ru

Cl 2 133 Ru

Cl 2 134 Ru

Cl 2 135 Ru

Cl 2 136 Ru

Cl 2 137 Ru

Cl 2 138 Ru

Cl 2 139 Ru

Cl 2 140 Ru

Cl 2 141 Ru

Cl 2 142 Ru

Cl 2 143 Ru

Cl 2 144 Ru

Cl 2 145 Ru

Cl 2 146 Ru

Cl 2 147 Ru

Cl 2 148 Ru

Cl 2 149 Ru

Cl 2 150 Ru

Cl 2 151 Ru

Cl 2 152 Ru

Cl 2 153 Ru

Cl 2 154 Ru

Cl 2 155 Ru

Cl 2 156 Ru

Cl 2 157 Ru

Cl 2 158 Ru

Cl 2 159 Ru

Cl 2 160 Ru

Cl 2 161 Ru

Cl 2 162 Ru

Cl 2 163 Ru

Cl 2 164 Ru

Cl 2 165 Ru

Cl 2 166 Ru

Cl 2 167 Ru

Cl 2 168 Ru

Cl 2 169 Ru

Cl 2 170 Ru

Cl 2 171 Ru

Cl 2 172 Ru

Cl 2 173 Ru

Cl 2 174 Ru

Cl 2 175 Ru

Cl 2 176 Ru

Cl 2 177 Ru

Cl 2 178 Ru

Cl 2 179 Ru

Cl 2 180 Ru

Cl 2 181 Ru

Cl 2 182 Ru

Cl 2 183 Ru

Cl 2 184 Ru

Cl 2 185 Ru

Cl 2 186 Ru

Cl 2 187 Ru

Cl 2 188 Ru

Cl 2 189 Ru

Cl 2 190 Ru

Cl 2 191 Ru

Cl 2 192 Ru

Cl 2 193 Ru

PF₆ 2 194 Ru

PF₆ 2 195 Ru

PF₆ 2 196 Ru

PF₆ 2 197 Ru

PF₆ 2 198 Ru

PF₆ 2 199 Ru

PF₆ 2 200 Ru

PF₆ 2 201 Ru

PF₆ 2 202 Ru

PF₆ 2 203 Ru

PF₆ 2 204 Ru

PF₆ 2 205 Ru

PF₆ 2 206 Ru

PF₆ 2 207 Ru

PF₆ 2 208 Ru

PF₆ 2 209 Ru

PF₆ 2 210 Ru

PF₆ 2 211 Ru

PF₆ 2 212 Ru

PF₆ 2 213 Ru

PF₆ 2 214 Ru

PF₆ 2 215 Ru

PF₆ 2 216 Ru

PF₆ 2 217 Ru

PF₆ 2 218 Ru

PF₆ 2 219 Ru

PF₆ 2 220 Ru

PF₆ 2 221 Ru

PF₆ 2 222 Ru

PF₆ 2 223 Ru

PF₆ 2 224 Ru

PF₆ 2 225 Ru

PF₆ 2 226 Ru

PF₆ 2 227 Ru

PF₆ 2 228 Ru

PF₆ 2 229 Ru

PF₆ 2 230 Ru

PF₆ 2 231 Ru

PF₆ 2 232 Ru

PF₆ 2 233 Ru

PF₆ 2 234 Ru

PF₆ 2 235 Ru

PF₆ 2 236 Ru

PF₆ 2 237 Ru

PF₆ 2 238 Ru

PF₆ 2 239 Ru

PF₆ 2 240 Ru

PF₆ 2 241 Ru

PF₆ 2 242 Ru

PF₆ 2 243 Ru

PF₆ 2 244 Ru

PF₆ 2 245 Ru

PF₆ 2 246 Ru

PF₆ 2 247 Ru

PF₆ 2 248 Ru

PF₆ 2 249 Ru

PF₆ 2 250 Ru

PF₆ 2 251 Ru

PF₆ 2 252 Ru

PF₆ 2 253 Ru

PF₆ 2 254 Ru

PF₆ 2 255 Ru

PF₆ 2 256 Ru

PF₆ 2 257 Ru

PF₆ 2 258 Ru

PF₆ 2 259 Ru

PF₆ 2 260 Ru

PF₆ 2 261 Ru

PF₆ 2 262 Ru

PF₆ 2 263 Ru

PF₆ 2 264 Ru

PF₆ 2 265 Ru

PF₆ 2 266 Ru

PF₆ 2 267 Ru

PF₆ 2 268 Ru

PF₆ 2 269 Ru

PF₆ 2 270 Ru

PF₆ 2 271 Ru

PF₆ 2 272 Ru

PF₆ 2 273 Ru

PF₆ 2 274 Ru

PF₆ 2 275 Ru

PF₆ 2 276 Ru

PF₆ 2 277 Ru

PF₆ 2 278 Ru

PF₆ 2 279 Ru

PF₆ 2 280 Ru

PF₆ 2 281 Ru

PF₆ 2 282 Ru

PF₆ 2 283 Ru

PF₆ 2 284 Ru

PF₆ 2 285 Ru

PF₆ 2 286 Ru

PF₆ 2 287 Ru

PF₆ 2 288 Ru

PF₆ 2 289 Ru

PF₆ 2 290 Ru

PF₆ 2 291 Ru

PF₆ 2 292 Ru

PF₆ 2 293 Ru

PF₆ 2 294 Ru

PF₆ 2 295 Ru

PF₆ 2 296 Ru

PF₆ 2 297 Ru

PF₆ 2 298 Ru

PF₆ 2 299 Ru

PF₆ 2 300 Ru

PF₆ 2 301 Ru

PF₆ 2 302 Ru

PF₆ 2 303 Ru

PF₆ 2 304 Ru

PF₆ 2 305 Ru

PF₆ 2 306 Ru

PF₆ 2 307 Ru

PF₆ 2 308 Ru

PF₆ 2 309 Ru

PF₆ 2 310 Ru

PF₆ 2 311 Ru

PF₆ 2 312 Ru

PF₆ 2 313 Ru

PF₆ 2 314 Ru

PF₆ 2 315 Ru

PF₆ 2 316 Ru

PF₆ 2 317 Ru

PF₆ 2 318 Ru

PF₆ 2 319 Ru

PF₆ 2 320 Ru

PF₆ 2 321 Ru

PF₆ 2 322 Ru

PF₆ 2 323 Ru

PF₆ 2 324 Ru

PF₆ 2 325 Ru

PF₆ 2 326 Ru

PF₆ 2 327 Ru

PF₆ 2 328 Ru

PF₆ 2 329 Ru

PF₆ 2 330 Ru

PF₆ 2 331 Ru

PF₆ 2 332 Ru

PF₆ 2 333 Ru

PF₆ 2 334 Ru

PF₆ 2 335 Ru

PF₆ 2 336 Ru

PF₆ 2 337 Ru

PF₆ 2 338 Ru

PF₆ 2 339 Ru

PF₆ 2 340 Ru

PF₆ 2 341 Ru

PF₆ 2 342 Ru

PF₆ 2 343 Ru

PF₆ 2 344 Ru

PF₆ 2 345 Ru

PF₆ 2 346 Ru

PF₆ 2 347 Ru

PF₆ 2 348 Ru

PF₆ 2 349 Ru

PF₆ 2 350 Ru

PF₆ 2 351 Ru

PF₆ 2 352 Ru

PF₆ 2 353 Ru

PF₆ 2 354 Ru

PF₆ 2 355 Ru

PF₆ 2 356 Ru

PF₆ 2 357 Ru

PF₆ 2 358 Ru

PF₆ 2 359 Ru

PF₆ 2 360 Ru

PF₆ 2 361 Ru

PF₆ 2 362 Ru

PF₆ 2 363 Ru

PF₆ 2 364 Ru

PF₆ 2 365 Ru

PF₆ 2 366 Ru

PF₆ 2 367 Ru

PF₆ 2 368 Ru

PF₆ 2 369 Ru

PF₆ 2 370 Ru

PF₆ 2 371 Ru

PF₆ 2 372 Ru

PF₆ 2 373 Ru

PF₆ 2 374 Ru

PF₆ 2 375 Ru

PF₆ 2 376 Ru

PF₆ 2 377 Ru

PF₆ 2 378 Ru

PF₆ 2 379 Ru

PF₆ 2 380 Ru

PF₆ 2 381 Ru

PF₆ 2 382 Ru

PF₆ 2 383 Ru

PF₆ 2 384 Ru

PF₆ 2

Exemplary embodiments include compounds having the formula (IX) or apharmaceutically acceptable salt form thereof:

wherein non-limiting examples of M, Lig, t, X, and q are defined hereinbelow in Table 4.

TABLE 4 Entry M Lig  t  X  q  1 Ru

1 Cl 4 2 Ru

2 Cl 4 3 Ru

3 Cl 4 4 Ru

4 Cl 4 5 Ru

5 Cl 4 6 Ru

6 Cl 4 7 Ru

1 PF₆ 4 8 Ru

2 PF₆ 4 9 Ru

3 PF₆ 4 10 Ru

4 PF₆ 4 11 Ru

5 PF₆ 4 12 Ru

6 PF₆ 4 13 Ru

1 Cl 4 14 Ru

2 Cl 4 15 Ru

3 Cl 4 16 Ru

4 Cl 4 17 Ru

5 Cl 4 18 Ru

6 Cl 4 19 Ru

1 PF₆ 4 20 Ru

2 PF₆ 4 21 Ru

3 PF₆ 4 22 Ru

4 PF₆ 4 23 Ru

5 PF₆ 4 24 Ru

6 PF₆ 4 25 Ru

1 Cl 4 26 Ru

2 Cl 4 27 Ru

3 Cl 4 28 Ru

4 Cl 4 29 Ru

5 Cl 4 30 Ru

6 Cl 4 31 Ru

1 PF₆ 4 32 Ru

2 PF₆ 4 33 Ru

3 PF₆ 4 34 Ru

4 PF₆ 4 35 Ru

5 PF₆ 4 36 Ru

6 PF₆ 4 37 Ru

1 Cl 4 38 Ru

2 Cl 4 39 Ru

3 Cl 4 40 Ru

4 Cl 4 41 Ru

5 Cl 4 42 Ru

6 Cl 4 43 Ru

1 PF₆ 4 44 Ru

2 PF₆ 4 45 Ru

3 PF₆ 4 46 Ru

4 PF₆ 4 47 Ru

5 PF₆ 4 48 Ru

6 PF₆ 4 49 Ru

1 Cl 4 50 Ru

2 Cl 4 51 Ru

3 Cl 4 52 Ru

4 Cl 4 53 Ru

5 Cl 4 54 Ru

6 Cl 4 55 Ru

1 PF₆ 4 56 Ru

2 PF₆ 4 57 Ru

3 PF₆ 4 58 Ru

4 PF₆ 4 59 Ru

5 PF₆ 4 60 Ru

6 PF₆ 4 61 Ru

1 Cl 4 62 Ru

2 Cl 4 63 Ru

3 Cl 4 64 Ru

4 Cl 4 65 Ru

5 Cl 4 66 Ru

6 Cl 4 67 Ru

1 PF₆ 4 68 Ru

2 PF₆ 4 69 Ru

3 PF₆ 4 70 Ru

4 PF₆ 4 71 Ru

5 PF₆ 4 72 Ru

6 PF₆ 4 73 Ru

1 Cl 4 74 Ru

2 Cl 4 75 Ru

3 Cl 4 76 Ru

4 Cl 4 77 Ru

5 Cl 4 78 Ru

6 Cl 4 79 Ru

1 PF₆ 4 80 Ru

2 PF₆ 4 81 Ru

3 PF₆ 4 82 Ru

4 PF₆ 4 83 Ru

5 PF₆ 4 84 Ru

6 PF₆ 4 85 Ru

1 Cl 4 86 Ru

2 Cl 4 87 Ru

3 Cl 4 88 Ru

4 Cl 4 89 Ru

5 Cl 4 90 Ru

6 Cl 4 91 Ru

1 PF₆ 4 92 Ru

2 PF₆ 4 93 Ru

3 PF₆ 4 94 Ru

4 PF₆ 4 95 Ru

5 PF₆ 4 96 Ru

6 PF₆ 4 97 Ru

1 Cl 4 98 Ru

2 Cl 4 99 Ru

3 Cl 4 100 Ru

4 Cl 4 101 Ru

5 Cl 4 102 Ru

6 Cl 4 103 Ru

1 PF₆ 4 104 Ru

2 PF₆ 4 105 Ru

3 PF₆ 4 106 Ru

4 PF₆ 4 107 Ru

5 PF₆ 4 108 Ru

6 PF₆ 4 109 Ru

1 Cl 4 110 Ru

2 Cl 4 111 Ru

3 Cl 4 112 Ru

4 Cl 4 113 Ru

5 Cl 4 114 Ru

6 Cl 4 115 Ru

1 PF₆ 4 116 Ru

2 PF₆ 4 117 Ru

3 PF₆ 4 118 Ru

4 PF₆ 4 119 Ru

5 PF₆ 4 120 Ru

6 PF₆ 4 121 Ru

1 Cl 4 122 Ru

2 Cl 4 123 Ru

3 Cl 4 124 Ru

4 Cl 4 125 Ru

5 Cl 4 126 Ru

6 Cl 4 127 Ru

1 PF₆ 4 128 Ru

2 PF₆ 4 129 Ru

3 PF₆ 4 130 Ru

4 PF₆ 4 131 Ru

5 PF₆ 4 132 Ru

6 PF₆ 4 133 Ru

1 Cl 4 134 Ru

2 Cl 4 135 Ru

3 Cl 4 136 Ru

4 Cl 4 137 Ru

5 Cl 4 138 Ru

6 Cl 4 139 Ru

1 PF₆ 4 140 Ru

2 PF₆ 4 141 Ru

3 PF₆ 4 142 Ru

4 PF₆ 4 143 Ru

5 PF₆ 4 144 Ru

6 PF₆ 4 145 Ru

1 Cl 4 146 Ru

2 Cl 4 147 Ru

3 Cl 4 148 Ru

4 Cl 4 149 Ru

5 Cl 4 150 Ru

6 Cl 4 151 Ru

1 PF₆ 4 152 Ru

2 PF₆ 4 153 Ru

3 PF₆ 4 154 Ru

4 PF₆ 4 155 Ru

5 PF₆ 4 156 Ru

6 PF₆ 4 157 Ru

1 Cl 4 158 Ru

2 Cl 4 159 Ru

3 Cl 4 160 Ru

4 Cl 4 161 Ru

5 Cl 4 162 Ru

6 Cl 4 163 Ru

1 PF₆ 4 164 Ru

2 PF₆ 4 165 Ru

3 PF₆ 4 166 Ru

4 PF₆ 4 167 Ru

5 PF₆ 4 168 Ru

6 PF₆ 4 169 Ru

1 Cl 4 170 Ru

2 Cl 4 171 Ru

3 Cl 4 172 Ru

4 Cl 4 173 Ru

5 Cl 4 174 Ru

6 Cl 4 175 Ru

1 PF₆ 4 176 Ru

2 PF₆ 4 177 Ru

3 PF₆ 4 178 Ru

4 PF₆ 4 179 Ru

5 PF₆ 4 180 Ru

6 PF₆ 4 181 Ru

1 Cl 4 182 Ru

2 Cl 4 183 Ru

3 Cl 4 184 Ru

4 Cl 4 185 Ru

5 Cl 4 186 Ru

6 Cl 4 187 Ru

1 PF₆ 4 188 Ru

2 PF₆ 4 189 Ru

3 PF₆ 4 190 Ru

4 PF₆ 4 191 Ru

5 PF₆ 4 192 Ru

6 PF₆ 4 193 Ru

1 Cl 4 194 Ru

2 Cl 4 195 Ru

3 Cl 4 196 Ru

4 Cl 4 197 Ru

5 Cl 4 198 Ru

6 Cl 4 199 Ru

1 PF₆ 4 200 Ru

2 PF₆ 4 201 Ru

3 PF₆ 4 202 Ru

4 PF₆ 4 203 Ru

5 PF₆ 4 204 Ru

6 PF₆ 4

Exemplary embodiments include compounds having the formula (X) or apharmaceutically acceptable salt form thereof:

wherein non-limiting examples of M, Lig, t, X, and q are defined hereinbelow in Table 5.

TABLE 5 Entry M Lig  t  X  q  1 Ru

1 Cl 4 2 Ru

2 Cl 4 3 Ru

3 Cl 4 4 Ru

4 Cl 4 5 Ru

5 Cl 4 6 Ru

6 Cl 4 7 Ru

1 PF₆ 4 8 Ru

2 PF₆ 4 9 Ru

3 PF₆ 4 10 Ru

4 PF₆ 4 11 Ru

5 PF₆ 4 12 Ru

6 PF₆ 4 13 Ru

1 Cl 4 14 Ru

2 Cl 4 15 Ru

3 Cl 4 16 Ru

4 Cl 4 17 Ru

5 Cl 4 18 Ru

6 Cl 4 19 Ru

1 PF₆ 4 20 Ru

2 PF₆ 4 21 Ru

3 PF₆ 4 22 Ru

4 PF₆ 4 23 Ru

5 PF₆ 4 24 Ru

6 PF₆ 4 25 Ru

1 Cl 4 26 Ru

2 Cl 4 27 Ru

3 Cl 4 28 Ru

4 Cl 4 29 Ru

5 Cl 4 30 Ru

6 Cl 4 31 Ru

1 PF₆ 4 32 Ru

2 PF₆ 4 33 Ru

3 PF₆ 4 34 Ru

4 PF₆ 4 35 Ru

5 PF₆ 4 36 Ru

6 PF₆ 4 37 Ru

1 Cl 4 38 Ru

2 Cl 4 39 Ru

3 Cl 4 40 Ru

4 Cl 4 41 Ru

5 Cl 4 42 Ru

6 Cl 4 43 Ru

1 PF₆ 4 44 Ru

2 PF₆ 4 45 Ru

3 PF₆ 4 46 Ru

4 PF₆ 4 47 Ru

5 PF₆ 4 48 Ru

6 PF₆ 4 49 Ru

1 Cl 4 50 Ru

2 Cl 4 51 Ru

3 Cl 4 52 Ru

4 Cl 4 53 Ru

5 Cl 4 54 Ru

6 Cl 4 55 Ru

1 PF₆ 4 56 Ru

2 PF₆ 4 57 Ru

3 PF₆ 4 58 Ru

4 PF₆ 4 59 Ru

5 PF₆ 4 60 Ru

6 PF₆ 4 61 Ru

1 Cl 4 62 Ru

2 Cl 4 63 Ru

3 Cl 4 64 Ru

4 Cl 4 65 Ru

5 Cl 4 66 Ru

6 Cl 4 67 Ru

1 PF₆ 4 68 Ru

2 PF₆ 4 69 Ru

3 PF₆ 4 70 Ru

4 PF₆ 4 71 Ru

5 PF₆ 4 72 Ru

6 PF₆ 4 73 Ru

1 Cl 4 74 Ru

2 Cl 4 75 Ru

3 Cl 4 76 Ru

4 Cl 4 77 Ru

5 Cl 4 78 Ru

6 Cl 4 79 Ru

1 PF₆ 4 80 Ru

2 PF₆ 4 81 Ru

3 PF₆ 4 82 Ru

4 PF₆ 4 83 Ru

5 PF₆ 4 84 Ru

6 PF₆ 4 85 Ru

1 Cl 4 86 Ru

2 Cl 4 87 Ru

3 Cl 4 88 Ru

4 Cl 4 89 Ru

5 Cl 4 90 Ru

6 Cl 4 91 Ru

1 PF₆ 4 92 Ru

2 PF₆ 4 93 Ru

3 PF₆ 4 94 Ru

4 PF₆ 4 95 Ru

5 PF₆ 4 96 Ru

6 PF₆ 4 97 Ru

1 Cl 4 98 Ru

2 Cl 4 99 Ru

3 Cl 4 100 Ru

4 Cl 4 101 Ru

5 Cl 4 102 Ru

6 Cl 4 103 Ru

1 PF₆ 4 104 Ru

2 PF₆ 4 105 Ru

3 PF₆ 4 106 Ru

4 PF₆ 4 107 Ru

5 PF₆ 4 108 Ru

6 PF₆ 4 109 Ru

1 Cl 4 110 Ru

2 Cl 4 111 Ru

3 Cl 4 112 Ru

4 Cl 4 113 Ru

5 Cl 4 114 Ru

6 Cl 4 115 Ru

1 PF₆ 4 116 Ru

2 PF₆ 4 117 Ru

3 PF₆ 4 118 Ru

4 PF₆ 4 119 Ru

5 PF₆ 4 120 Ru

6 PF₆ 4 121 Ru

1 Cl 4 122 Ru

2 Cl 4 123 Ru

3 Cl 4 124 Ru

4 Cl 4 125 Ru

5 Cl 4 126 Ru

6 Cl 4 127 Ru

1 PF₆ 4 128 Ru

2 PF₆ 4 129 Ru

3 PF₆ 4 130 Ru

4 PF₆ 4 131 Ru

5 PF₆ 4 132 Ru

6 PF₆ 4 133 Ru

1 Cl 4 134 Ru

2 Cl 4 135 Ru

3 Cl 4 136 Ru

4 Cl 4 137 Ru

5 Cl 4 138 Ru

6 Cl 4 139 Ru

1 PF₆ 4 140 Ru

2 PF₆ 4 141 Ru

3 PF₆ 4 142 Ru

4 PF₆ 4 143 Ru

5 PF₆ 4 144 Ru

6 PF₆ 4 145 Ru

1 Cl 4 146 Ru

2 Cl 4 147 Ru

3 Cl 4 148 Ru

4 Cl 4 149 Ru

5 Cl 4 150 Ru

6 Cl 4 151 Ru

1 PF₆ 4 152 Ru

2 PF₆ 4 153 Ru

3 PF₆ 4 154 Ru

4 PF₆ 4 155 Ru

5 PF₆ 4 156 Ru

6 PF₆ 4 157 Ru

1 Cl 4 158 Ru

2 Cl 4 159 Ru

3 Cl 4 160 Ru

4 Cl 4 161 Ru

5 Cl 4 162 Ru

6 Cl 4 163 Ru

1 PF₆ 4 164 Ru

2 PF₆ 4 165 Ru

3 PF₆ 4 166 Ru

4 PF₆ 4 167 Ru

5 PF₆ 4 168 Ru

6 PF₆ 4 169 Ru

1 Cl 4 170 Ru

2 Cl 4 171 Ru

3 Cl 4 172 Ru

4 Cl 4 173 Ru

5 Cl 4 174 Ru

6 Cl 4 175 Ru

1 PF₆ 4 176 Ru

2 PF₆ 4 177 Ru

3 PF₆ 4 178 Ru

4 PF₆ 4 179 Ru

5 PF₆ 4 180 Ru

6 PF₆ 4 181 Ru

1 Cl 4 182 Ru

2 Cl 4 183 Ru

3 Cl 4 184 Ru

4 Cl 4 185 Ru

5 Cl 4 186 Ru

6 Cl 4 187 Ru

1 PF₆ 4 188 Ru

2 PF₆ 4 189 Ru

3 PF₆ 4 190 Ru

4 PF₆ 4 191 Ru

5 PF₆ 4 192 Ru

6 PF₆ 4 193 Ru

1 Cl 4 194 Ru

2 Cl 4 195 Ru

3 Cl 4 196 Ru

4 Cl 4 197 Ru

5 Cl 4 198 Ru

6 Cl 4 199 Ru

1 PF₆ 4 200 Ru

2 PF₆ 4 201 Ru

3 PF₆ 4 202 Ru

4 PF₆ 4 203 Ru

5 PF₆ 4 204 Ru

6 PF₆ 4

For the purposes of the present invention, a compound depicted by theracemic formula will stand equally well for either of the twoenantiomers or mixtures thereof, or in the case where a second chiralcenter is present, all diastereomers.

In all of the embodiments provided herein, examples of suitable optionalsubstituents are not intended to limit the scope of the claimedinvention. The compounds of the invention may contain any of thesubstituents, or combinations of substituents, provided herein.

Photophysical properties of the compounds of the invention:

The modular nature of compounds of the disclosure enables fine-tuning oftheir photophysical properties through minor changes to the molecularscaffold. For example, in the series 1a, 10a, and 14a (FIG. 2), theabsorption spectra systematically shift to longer wavelengths, leadingto corresponding increases in the extinction coefficients in the visibleregion (Table 6). When R is a single thiophene as in 1a, the integratedabsorption of visible light (ε vs. cm⁻¹) is approximately 5841 (FIG. 2).Incorporation of additional thiophene units in the pendant R groupincreases this absorption cross-section; for 10a and 14a, theseenhancements are 40% and 141%, respectively (Table 7). According to thisprogression, the absorption of visible light will be increasesystematically in the corresponding homoleptic complexes as the numberof thiophene linkers is increased. Such improvements in visible lightabsorption are critical to the optimization of PDCs for applicationssuch as PDT.

TABLE 6 Electronic absorption and emission maxima for PDCs and thereference compounds Complex λmax Absorption (log ε), nm λ_(em), nm  1a242 (4.54), 286 (5.00), 326 (4.42), 426 (4.10), 458 616 (4.21), 500(3.37)  1b 220 (4.91), 260 (5.04), 286 (4.83), 326 (4.37), 424 604(4.25), 454 (4.31), 500 (3.47)  2a 244 (4.90), 250 (4.91), 284 (5.20),378 (4.76), 396 617 (4.79), 430 (4.66), 460 (4.70), 500 (3.89)  2b 220(5.15), 260 (5.29), 284 (4.81), 298 (4.63), 374 605 (4.70), 396 (4.78),458 (4.63), 500 (3.86)  3a 252 (4.86), 282 (5.13), 422 (4.79), 464(4.75) 500 619 (3.94)  3b 220 (4.96), 260 (5.10), 284 (4.59), 298(4.29), 420 613 (4.61), 462 (4.55), 500 (3.87) 10a 252 (4.59), 284(4.87), 370 (4.60), 458 (4.28), 500 618 (3.46) 10b 220 (4.95), 260(5.07), 286 (4.58), 294 (4.52) 378 618 (4.69), 456 (4.41), 500 (3.62)14a 252 (4.61), 282 (4.83), 408 (4.62), 464 (4.37), 500 620 (3.60) 14b220 (4.81), 260 (4.92), 286 (4.45), 410 (4.59) 458 618 (4.37), 500(3.53)

TABLE 7 Integrated absorption profiles for PDCs in the visible range(25,000-15,000 cm⁻¹). Complex Abs. (400-700 nm) increase  1a 5841 10a8190  40% (1.4X) 14a 14,100 141% (2.4X)

The modular nature of the compounds of the disclosure enablesfine-tuning of their photophysical properties through systematic changesto the molecular scaffold. For example, In the series 1a, 10a, 14a, and16a, the absorption spectra extend to longer wavelengths as the numberof thiophene rings is increased, leading to absorption in the nearinfrared region when 4 thiophenes are present (16a). Also, theintegrated absorption from 400-900 nm is almost 4 times greater for 16aversus 1a (Table 8 and FIG. 3).

TABLE 8 Integrated absorption profiles for PDCs 1a, 10a, 14a, and 16a inthe visible and NIR region. Integrated absorption relative Compoundincrease (400-900 nm) Increase relative to 1a  1a 19 1 10a 33 1.74 14a50 2.63 16a 69 3.63

Current sensitizers for PDT rely on ¹O₂ generation for photodynamicaction toward target cells. Similar to the trend in visible absorption,the quantum yields for ¹O₂ production for these compounds also increasewith the number of thiophene units (FIG. 4). The measured Φ¹O₂ for 1a is0.47 while 10a and 14a yield 0.74 and 1.0, respectively (Table 9). Twoadditional compounds in this family including 14a (FIGS. 1, 3a and 3b)have been shown to exhibit 100% efficiency for ¹O₂ sensitization,exceeding the PDT drug PHOTOFRIN (Φ¹O₂=0.75, ethanol). Interestingly,despite remarkable ¹O₂ quantum yields, compounds of the disclosure havea quantum yield for emission less than 1% in deoxygenated solution (e.g.14a, Φ¹O₂<0.15%). Therefore, another nonradiative pathway becomesimportant in the absence of O₂ and may predominate under hypoxicconditions, imparting O₂ independence to the compounds of the disclosureas a PDT agent. The fact that compounds of the disclosure photodamageDNA under hypoxic conditions is evidence for potential O₂ independencein terms of photobiological activity.

TABLE 9 Photophysical properties for PDCs τ_(em), ns τ_(em), ns ComplexΦ_(em) (air) Φ_(em) (Ar)* (air) (Ar)* Φ¹O₂  1a 1.27 × 10⁻² 1.08 × 10⁻¹150 1040 0.467  1b 8.92 × 10⁻³ 7.40 × 10⁻² 112 700 0.422  2a 4.02 × 10⁻³5.51 × 10⁻² 188 2810 0.741  2b 2.15 × 10⁻³ 5.19 × 10⁻² 194 4400 0.677 3a 3.60 × 10⁻⁴ 1.96 × 10⁻³ 159 1520 1  3b 5.40 × 10⁻⁴ 2.06 × 10⁻³ 1291240 1 10a 4.59 × 10⁻⁴ 3.37 × 10⁻² 151 1290 0.738 10b 3.83 × 10⁻⁴ 1.14 ×10⁻² 133 741 0.790 14a 3.30 × 10⁻⁴ 1.34 × 10⁻³ 154 575 1 14b 4.70 × 10⁻⁴1.88 × 10⁻³ 140 908 0.496 *Argon purge, 30 min. at 30 ± 5 mm Hg.

DNA Binding Properties of the Compounds of the Disclosure:

As shown in FIG. 5, 10a binds DNA very strongly, having one of thelargest known binding constants for this type of interaction(K_(b)=9.1×10⁷ at 372 nm; 4.4×10⁷ at 460 nm) determined by UV-Visabsorption. The magnitude of this binding interaction is furthersupported by emission measurements (K_(b)=4.2×10⁷ at 625 nm) and is alsoobserved for other PDCs of the disclosure such as 14a. This bindingoccurs in cells and can be seen as diffuse nuclear staining by the PDC14a in HL60 cells when viewed by laser scanning confocal microscopy(FIG. 6).

Changes in the identity of the ancillary ligands (10b) or a reduction inthe number of thiophenes in the C2-substituted ligand (1a) attenuatesthe binding affinity by an order of magnitude, indicating that subtlemodifications lead to profound effects within a single family ofstructurally related PDCs of the disclosure (Table 10).

TABLE 10 DNA binding constants from absorption and emission opticaltitrations Complex K_(b), s (Abs.) K_(b), s (Em.)  1a 2.4 × 10⁶, 0.73(456 nm) 3.2 × 10⁶, 4.7 (627 nm) 10a 4.4 × 10⁷, 0.4 (460 nm)  4.2 × 10⁷,7.6 (625 nm) 10b 3.3 × 10⁶, 0.19 (462 nm) 3.6 × 10⁷, 7.1 (619 nm) 14a1.6 × 10⁷, 0.30 (452 nm) 1.4 × 10⁷, 5.1 (626 nm)

The PDCs of the disclosure also differ in the extent to which theystabilize a DNA helix when they bind (FIG. 7, Table 11). Even though 1aexhibits weaker binding to DNA relative to 10a, the additional thiophenemoiety in 10a leads to less stabilization of the helix upon DNA bindingas observed by only a slight increase in T_(m) of 1° C. In fact, gelelectrophoretic analysis indicates that 10a substantially destabilizesthe native DNA structure, evidenced by the inability of the intercalatorethidium bromide to stain DNA effectively at [10a]: [DNA]>0.4 (FIG. 14,(b) Lanes 8-14); in contrast, 1a does not interfere with ethidiumbromide intercalation, and the gel bands produced by ethidium bromidestaining remain visible with increasing PDC concentration (FIG. 14, (a)Lanes 8-14).

TABLE 11 Thermal denaturation parameters for CT-DNA with added 1a, 1b,10a, and 10b ([PDC]/[NP] = 0.1). T_(m) for CT-DNA in the absence of aPDC was 78.6° C. Complex T_(m),° C. ΔT_(m)  1a 82.7 4.1  1b 86.0 7.4 10a79.9 1.3 10b 79.9 1.3

DNA Light Switch Effects of the Compounds of the Invention

While the PDCs of the disclosure, excluding 1a and 1b, have a quantumyield for emission of less than 1% in air, they become measurablyemissive in the absence of oxygen (Table 9) or in the presence of DNA(FIG. 8), the latter of which is known as the DNA light-switch effect.This effect increases with the number of thiophene units in the pendantR group: approximately 2-, 3-, and 4-fold for 1a, 10a, and 14a,respectively (Table 12).

TABLE 12 Measured enhancement for DNA light-switch effect produced byPDCs 1a, 10a, and 14a Complex Enhancement  1a  76% (1.8x) 10a 160%(2.6x) 14a 260% (3.6x)

The enhancement in luminescence can be exploited to detect the PDCs ofthe disclosure in the presence of biomolecules and/or hypoxicenvironments or to ascertain subcellular localization. For example, 10alocalizes outside of the nucleus in non-irradiated cells (FIG. 10 c)while 14a appears to stain chromatin (FIG. 12 c). In irradiated cells,both 10a and 14a are distributed throughout the cytoplasm and nucleus(FIGS. 10 d and 12 d). Interestingly, 10a and 10b stain nonviable cellsin the dark at 5 minutes (FIGS. 10 b and 11 b). 1a, being 1-2 orders ofmagnitude more emissive than 10a or 14a in air or argon, can be used asa stain at much lower concentration.

DNA photo cleavage properties of the compounds of the disclosure:

Perhaps due, in part, to very strong binding interactions with DNA, PDCsof the disclosure cause extensive DNA damage in the form ofsingle-strand breaks upon irradiation with visible light. Theobservation that no strand breaks occur at similar concentrations of PDCwithout light activation is very promising as a mechanism for cellulardestruction based on photodynamic action (FIGS. 13 and 14). Given thatDNA is the blueprint for all cellular function, photo dynamic actiontargeted at this biomolecule will initiate apoptotic pathways thatselectively destroy irradiated cells that have taken up the PDC of thedisclosure. The mechanism for DNA damage is not confirmed, but with ¹O₂quantum yields of unity for PDCs such as 14a, it is likely that¹O₂-mediated photodamage plays a significant role in oxygenatedconditions. Under hypoxic conditions, DNA photodamage by these compoundsremains significant (FIGS. 15 and 16, Tables 12 and 13), indicating thatoxygen-deficient cells are susceptible to photodynamic action by PDCssuch as 10a and 14a. This oxygen-independent DNA damage is furthercorroborated by the fact that the quantum yield for emission for acompound such as 14a remains very low (0.13%) in the absence of oxygen.Therefore, intra- or intermolecular electron transfer, which is commonto thiophene photophysics, contribute to non-radiative decay in thesesystems as well. Such electron transfer pathways may become important inthe absence of molecular oxygen, giving rise to Type I photoprocesseswith biomolecules. Preliminary experiments indicate that energy versuselectron transfer is governed by environment. In other words, there maynot be a partioning of excited state energy between numerous pathways;rather, the presence of oxygen serves to switch between two majornon-radiative pathways: electron transfer and energy transfer, both ofwhich are extremely efficient at invoking DNA damage. This photodynamicswitch between Type I and Type II photoprocesses ensures thatphotodynamic action is optimal regardless of oxygen concentration. Sucha versatile PDC eliminates the need for distinct Type I and Type IIphotosensitizers for maximizing photodynamic action according to oxygenlevels in target cells.

TABLE 12 Calculated percentages of DNA forms observed for PDC- mediatedpUC19 photocleavage in air-saturated and deoxygenated solution with420-nm irradiation, FIG. 15. Lane PDC Treatment Form I Form II Form III1 None Air, dark 76 24 2 [Ru(bpy)³]²⁺ Air, dark 71 29 3 [Ru(bpy)³]²⁺Air, +hv  33 67 4 [Ru(bpy)³]²⁺ Ar, +hv 71 29 5 10a Air, dark 78 22 6 10aAir, +hv  14 86 7 10a Ar, +hv 34 66 8 10b Air, dark 71 29 9 10b Air,+hv  15 85 10 10b Ar, +hv 27 73 11 14a Air, dark 50 23 27 12 14a Air,+hv  21 79 13 14a Ar, +hv 30 39 31

TABLE 13 Calculated percentages of DNA forms observed for PDC- mediatedpUC19 photocleavage in air-saturated and deoxygenated solution withvisible irradiation, FIG. 16. Lane Complex Treatment Form I Form II 1None Air, dark 77 23 2 [Ru(bpy)³]²⁺ Air, dark 72 28 3 [Ru(bpy)³]²⁺ Air,+hv  37 63 4 [Ru(bpy)³]²⁺ Ar, +hv 57 43 5 10a Air, dark 80 20 6 10a Air,+hv  14 86 7 10a Ar, +hv 38 62

Cytotoxicity and Photocytotoxicity of the Compounds of the Invention

A host of PDCs of the disclosure exhibit marked photocytotoxicity withno substantial dark toxicity even at concentrations as high as 100 μM(checked at 12, 18, 24, 36, and 48 hr. post-irradiation). As observedfor 1a, 10a, and 14a in terms of integrated visible absorption, Φ¹O₂,DNA binding, and DNA photodamage, there exists a profound effect ofoverall structure on the photocytotoxicity toward HL-60 cells. Toillustrate, 10a, which differs from 10b only in thatLig=[2,2′]bipyridine instead of [1,10]phenanthroline, is phototoxictoward cells at 20 μM, whereas 10b displays no photocytotoxicity even at100 μM (FIG. 17). For the series 1a, 10a, and 14a, whereLig=[2,2′]bipyridine and R¹ differs by the number of thiophenes thatmake up the pendant C2 unit, there is a drastic difference in thephotocytotoxicity toward HL-60 cells, whereby photodynamic action is100% at less than 10 μM for 14a and virtually zero at similarconcentrations of 1a (FIG. 18). Interestingly, as the number ofthiophenes that make up R¹ increases, so does the photodynamic actiontoward HL-60 cells. This progression was confirmed through viabilitystaining with Trypan Blue using a commercial cell counting system aswell as AO-EB viability staining using epi-fluorescence microscopy (FIG.19). This activity parallels the trends observed for integrated visibleabsorption, Φ¹O₂, and DNA photodamage, which together determine theoverall photocytotoxicity for a given PDC toward a particular cell.

To discern whether batch-to-batch variability in photodynamic actionexists for the PDCs of the disclosure, 1a, 10a, and 14a were synthesizedusing a novel method for preparing compounds of the disclosure usingmicrowave irradiation. Their molecular structures were confirmed by ¹HNMR, HRMS, and elemental analysis and are identical to structuresobtained via standard methodology. The photocytotoxicity observed forthe PDCs 1a, 10a, and 14a prepared by microwave irradiation exhibits thesame trend that was documented for PDCs 1a, 10a, and 14a preparedaccording to standard methods: namely, 14a>10a>1a (FIG. 20). Likewise,the effective concentrations at which photocytotoxicity is observed foreach PDC does not vary substantially between the two independentlyprepared batches for the post-irradiation times employed (18 hr., FIG.21 and 40 hr., FIG. 22). The photodynamic action of 14a is bestillustrated in FIGS. 23-25, where 100% photocytotoxicity occurs in the5-10 μM range.

Activation of Apoptotic Pathways by Compounds of the Disclosure

Both 10a and 14a exert their photocytotoxicity by activating apoptoticpathways. Nuclear condensation and the formation of apoptotic bodies canbe seen clearly with AO-EB nuclear morphology staining at 18 and 40 hr.post-irradiation by epi-fluorescence microscopy of cells treated with10a (FIG. 26, Panel C). AO stains viable nuclei and fluoresces intensegreen (FIG. 26, Panel B, upper right cell) while EB stains nonviablenuclei and fluoresces intense red (FIG. 26, Panel C, lower left cell).Likewise nuclear condensation and the formation of apoptotic bodies areevident in AO-EB stained cells treated with 14a at 18 (FIG. 27) and 40hr. (FIG. 28) post-irradiation.

Further support for apoptosis as the primary mechanism for cellulardestruction is based on the large changes in cell diameter that takeplace upon irradiation of cells that have been treated with 10a or 14a.It is well-known that apoptotic cell diameters are significantly smallerthan either normal or necrotic cells. Healthy HL-60 cells can range from9-25 μm in diameter, although 13 μm is typical. The HL-60 cells used inthese cytotoxicity studies average 11-13 μm in diameter. In the presenceof 50 μM 10a, 70% of the irradiated HL-60 cells that remain at 40 hr.are roughly half their original size (mean diameter <6 μm) while thesame concentration of 10b has little effect on the mean cell diameter(FIG. 29). The same trend exists for 14a while 1a, which exhibits nosignificant photocytotoxicity, does not notably alter the irradiatedHL-60 cell diameter distribution (FIG. 30). Interestingly, 1a doesproduce a small population (<30% at 40 hr.) of HL-60 cells withdiameters less than 6 μm in the dark. This finding is corroborated bythe slight dark toxicity observed for 1a (FIG. 18 a). Together, thesechanges in size distribution upon irradiation of PDC treated cellsfurther underscore that the structure of compounds of the disclosure hasan important and distinct effect on the photobiology of these PDCs ofthe disclosure. The systematic variation that can be achieved in termsof their photophysical, photochemical, and photobiological propertiesoffers an avenue to rational PDC design and optimization of compounds ofthe disclosure.

PDCs of the Disclosure as Excited-State Oxidants and Reductants:

Compounds of the disclosure are capable of acting as excited-stateoxidants and reductants. For example, The excited-state reduction andoxidation potentials of 14a are estimated at 1.31 and −0.87 V,respectively, making both guanine oxidation and cystosine or thyminereduction feasible upon photoactivation (Table 14, FIGS. 31 and 32).Compounds of the disclosure act as photoreductants for DNA, supported bythe observation that endogenous reductants such as glutathione (GSH) andascorbic acid (AA) facilitate DNA photodamage by compounds of thedisclosure.

TABLE 14 Ground state and excited-state redox potentials of 14a EntryE_(ox)(V) E_(red)(V) E₀₋₀(eV) E*_(red)(V) E*^(ox)(V) 14a 1.56 −1.13 2.431.31 −0.87 −1.33 1.10 −1.68 0.75

The DNA photocleavage produced by 14a is greatly enhanced by endogenousreductants such as glutathione (GSH). At submicromolar concentrations of14a, 4 mM GSH results in detectable double-strand breaks while DNAexposed to the PDC alone is mostly undamaged (FIG. 33, Lane 1). At 3 μM14a, the presence of GSH results in total DNA degradation (FIG. 33 Lane6). It is well-known that GSH concentraion is high in human tumourcells, decreasing the cytotoxic effects of electrophilic alkylatingagents (such as cisplatin) and radiation and also diminishing theeffects of PDT. Herein we disclose a family of PDT agents that ispreferentially activated by GSH.

The ability of GSH to facilitate DNA damage is broad in scope and holdsfor additional compounds of the disclosure (e.g., 14c and 16c), and forother reductants (ascorbate, DTT, NADH, etc.). FIG. 34 (Lanes 7-16)illustrates that for 1a, the addition of GSH (0.5-7 mM) has no effect onDNA photocleavage, and the predominant form of DNA is supercoiled (FormI). For 10a, GSH promotes the formation of both single- anddouble-strand DNA breaks, and for 14a, GSH promotes total degradation ofthe DNA. For all 3 complexes, Lane 17 shows the amount of DNA damage topBR322 that results with photoactivation of 2 μM PDC in the absence ofGSH. For comparison, Lane 14 shows the effect of 6 mM GSH on DNAphotodamage by the PDCs: (a) there is no effect on DNA photocleavage by1a; (b) GSH facilitates the formation of double-strand breaks by 10a;and (c) GSH facilitates total DNA degradation by 14a. FIG. 35demonstrates the facilitation of DNA photodamage by 14a for anotherreductant, ascorbic acid (AA). A comparison between Lanes 13 and 14clearly shows that the presence of AA turns a predominantlysingle-strand DNA photocleaver into a very powerful DNA degrader (notesmear in Lanes 9-13).

Destruction of Cancer Cells by Compounds of the Invention

Compounds of the disclosure with low dark toxicity are able toeffectively destroy cancer cells by PDT at very low concentrations exvivo (i.e., in a cell outside of an animal). An ideal PDC should exhibitPDT effects at low PDC concentrations while having a low dark toxicity,i.e., the cell kill of the PDC alone with no light exposure. Darktoxicity and PDT effects on cell kill for PDCs 10A, 14A, and 16A weretested ex vivo on the following cell lines: CT26.WT (murine coloncarcinoma), U87 (human glioblastoma), F98 (rat glioma). Irradiation wasconducted at 45 J/cm² (TLC-3000 light source, λ=530 nm, 4-6 hoursPS-light interval) and cell viability was measured 24 hourspost-irradiation using the Presto Blue cell viability assay. FIG. 39shows that neither 10A, 14A, nor 16A alone (without light) demonstratedsignificant cell kill. However, a strong photodynamic effect (PDC pluslight) was observed with 10A, 14A, and 16A; 60-100% cell kill in allthree cell lines, depending on the PDC concentration. The best effect oncell kill was shown for 14A and 16A, followed by 10A.

In Vivo Efficacy of Compounds of the Disclosure:

Compounds of the disclosure have comparable effective in vivo PDT dosesas FDA approved PHOTOFRIN and Aminolevulinic Acid (ALA). The mean toxicdose that causes death in 50% of animals tested (MTD₅₀) was determinedfor PDCs 14A and 14C. MTD₅₀ was identified by administering a series ofincreasing and decreasing drug doses, starting at a concentration 10²lower in magnitude to the presumed MTD₅₀ from in vitro studies. MTD₅₀sfor PDCs 14A and 14C are the effective doses that are used in furtherPDT studies. As shown in FIG. 40, the effective PDT doses for 14A and14C fall between the range of PHOTOFRIN and ALA, i.e., 36 mg/kg for 14A,98 mg/kg for 14C, 10 mg/kg for PHOTOFRIN, and 250 mg/kg for ALA. Note:the effective dose of protoporphyrin IX (PPIX, the photoactivemetabolite of ALA), is ¼ the administrative dose of ALA.

Compounds of the Disclosure Ex Vivo Potency

Compounds of the disclosure are more effective at killing cancer cellsex vivo as compared to ALA. For example, 14A is more effective atkilling cancer cells ex vivo compared to ALA. ALA is an FDA approved PDCcurrently used to treat skin, bladder, and brain cancers. As shown inFIG. 41, dark toxicity of 14A (PDC alone and without light) was weak ornegligible in all three cancer cell lines (CT26.WT, U87, F980) which wascomparable to the level of dark toxicity of ALA at the sameconcentrations. The efficiency of 14A plus light (TLC-3000 light source,530 nm, 45 J/cm²) in killing cells was also much higher compared to theefficacy of ALA. 14A resulted in 100% cell kill at very lowconcentrations, whereas ALA produced no, or very little, cell kill.

Tissue Penetration of Compounds of the Disclosure

PDCs should absorb light at longer wavelengths in order to effectivelypenetrate tissue. Absorbance at shorter wavelengths will achieve lesstissue penetration and may lead to skin photosensitivity. To measure theabsorbance for each PDC, PDC stock solutions were diluted with the samesolvent to achieve an optical density (OD) of 0.2 at 525 nm (6.7 μM for14C, 1.2 μM for PPIX). Optical densities were measured from 300 nm to800 nm. As shown in FIG. 42, both 14C and PPIX have similar andrelatively low absorption at 525 nm. PDT irradiation at wavelengths of525 nm or longer, as seen with compounds of the disclosure, areclinically warranted, while the use of shorter wavelengths is notclinically justified.

Photostability of Compounds of the Disclosure

Compounds of the disclosure exhibit greater photostability compared toPPIX. An effective PDC should maintain high photochemical stability,allowing for a long shelf-life. To measure photostability, each PDC wasirradiated and the OD at the wavelengths corresponding to the maximalabsorbance (423 nm for 14C, 411 nm for PPIX), was measured every 5minutes for 60 minutes of total irradiation time. FIG. 43 shows that PDC14C is photobleached by approximately 50%. PPIX underwent considerablephotobleaching by 60 minutes of irradiation (approximately 70%). 14C is,therefore, more photostable than PPIX.

Will the subcutaneous tumor data be inserted here?

Process

The present invention further relates to a process for preparing thephotodynamic compounds of the present invention.

Compounds of the present teachings can be prepared in accordance withthe procedures outlined herein, from commercially available startingmaterials, compounds known in the literature, or readily preparedintermediates, by employing standard synthetic methods and proceduresknown to those skilled in the art. Standard synthetic methods andprocedures for the preparation of organic molecules and coordinationcomplexes and functional group transformations and manipulations can bereadily obtained from the relevant scientific literature or fromstandard textbooks in the field. It will be appreciated that wheretypical or preferred process conditions (i.e., reaction temperatures,times, mole ratios of reactants, solvents, pressures, etc.) are given,other process conditions can also be used unless otherwise stated.Optimum reaction conditions can vary with the particular reactants orsolvent used, but such conditions can be determined by one skilled inthe art by routine optimization procedures. Those skilled in the art oforganic and inorganic synthesis will recognize that the nature and orderof the synthetic steps presented can be varied for the purpose ofoptimizing the formation of the compounds described herein.

The processes described herein can be monitored according to anysuitable method known in the art. For example, product formation can bemonitored by spectroscopic means, such as nuclear magnetic resonancespectroscopy (e.g., 1H or 13C), infrared spectroscopy, spectrophotometry(e.g., UV-visible), mass spectrometry, or by chromatography such as highpressure liquid chromatography (HPLC), gas chromatography (GC),gel-permeation chromatography (GPC), or thin layer chromatography (TLC).

Preparation of the compounds can involve protection and deprotection ofvarious chemical groups. The need for protection and deprotection andthe selection of appropriate protecting groups can be readily determinedby one skilled in the art. The chemistry of protecting groups can befound, for example, in Greene et al., Protective Groups in OrganicSynthesis, 2d. Ed. (Wiley & Sons, 1991), the entire disclosure of whichis incorporated by reference herein for all purposes.

The reactions or the processes described herein can be carried out insuitable solvents which can be readily selected by one skilled in theart of organic and inorganic synthesis. Suitable solvents typically aresubstantially nonreactive with the reactants, intermediates, and/orproducts at the temperatures at which the reactions are carried out,i.e., temperatures that can range from the solvent's freezingtemperature to the solvent's boiling temperature. A given reaction canbe carried out in one solvent or a mixture of more than one solvent.Depending on the particular reaction step, suitable solvents for aparticular reaction step can be selected.

The compounds of these teachings can be prepared by methods known in theart of organic and inorganic chemistry. The reagents used in thepreparation of the compounds of these teachings can be eithercommercially obtained or can be prepared by standard proceduresdescribed in the literature. For example, compounds of the presentinvention can be prepared according to the method illustrated in theGeneral Synthetic Schemes:

General Synthetic Schemes for Preparation of Compounds

The reagents used in the preparation of the compounds of this inventioncan be either commercially obtained or can be prepared by standardprocedures described in the literature. In accordance with thisinvention, compounds in the genus may be produced by one of thefollowing reaction schemes.

Compounds of formula (I) may be prepared according to the processoutlined in the following schemes.

Accordingly, a suitably substituted compound of the formula (XI), aknown compound or compound prepared by known methods, is reacted with asuitably substituted compound of the formula (XII) in the presence of anammonium salt such as ammonium acetate, ammonium formate, ammoniumchloride, ammonium bromide, ammonium sulfate and the like in a solventsuch as acetic acid, formic acid, propionic acid and the like,optionally in the presence of methanol, ethanol, N,N-dimethylformamideand the like, optionally heated, optionally heated with microwaveirradiation, to provide a compound of the formula (XIII) A compound ofthe formula (XIII) was then reacted with a compound of the formula (XIV)in a solvent such as methanol, ethanol, isopropanol, ethylene glycol,glycerol, water, 1,4-dioxane, dimethyl formamide, mixtures of theaforementioned solvents, and the like, optionally heated, optionallyheated with microwave irradiation, to provide a compound of the formula(II). Compounds of the formula (II) may be converted to alternative saltforms by conventional methods known to those skilled in the art.

Compounds of formula (III) may be prepared according to the processoutlined in Scheme 2.

Accordingly, a suitably substituted compound of the formula (XI) isreacted with a compound of the formula (XV) in an a solvent such asmethanol, ethanol, isopropanol, ethylene glycol, glycerol, water,1,4-dioxane, dimethyl formamide, mixtures of the aforementionedsolvents, and the like, optionally heated, optionally heated withmicrowave irradiation, to provide a compound of the formula (III).Compounds of the formula (III) may be converted to alternative saltforms by conventional methods known to those skilled in the art.

Compounds of formula (IV) may be prepared according to the processoutlined in Scheme 3.

Accordingly, a suitably substituted compound of the formula (XI) isreacted with a compound of the formula (XVI) in a solvent such asmethanol, ethanol, isopropanol, ethylene glycol, glycerol, water,1,4-dioxane, dimethyl formamide, mixtures of the aforementionedsolvents, and the like, optionally heated, optionally heated withmicrowave irradiation, to provide a compound of the formula (IV).Compounds of the formula (IV) may be converted to alternative salt formsby conventional methods known to those skilled in the art.

Compounds of formula (V) may be prepared according to the processoutlined in Scheme 4.

Accordingly, a suitably substituted compound of the formula (XI), aknown compound or compound prepared by known methods, is reacted with asuitably substituted compound of the formula (XVI) in the presence of anammonium salt such as ammonium acetate, ammonium formate, ammoniumchloride, ammonium bromide, ammonium sulfate and the like in a solventsuch as acetic acid, formic acid, acid, propionic acid and the like,optionally in the presence methanol, ethanol, N,N-dimethylformamide andthe like, optionally heated, optionally heated with microwaveirradiation, to provide a compound of the formula (XVII). A compound ofthe formula (XVII) was then reacted with a compound of the formula(XVIII) in a solvent such as methanol, ethanol, isopropanol ethyleneglycol, glycerol, water, 1,4-dioxane and the like, optionally heated,optionally heated with microwave irradiation, to provide a compound ofthe formula (V). Compounds of the formula (V) may be converted toalternative salt forms by conventional methods known to those skilled inthe art.

The invention will be illustrated in more detail with reference to thefollowing Examples, but it should be understood that the presentinvention is not deemed to be limited thereto.

The Examples provided below provide representative methods for preparingexemplary compounds of the present invention. The skilled practitionerwill know how to substitute the appropriate reagents, starting materialsand purification methods known to those skilled in the art, in order toprepare the compounds of the present invention.

¹H-NMR spectra were obtained on a Bruker Avance 300-MHz NMR. Low andhigh resolution mass spectral data were determined with a BrukerDaltonics micrOTOF instrument.

EXAMPLES

Preparation of 14L: 10-Phenanthroline-5,6-dione (166.6 mg, 0.800 mmol),ammonium acetate (616 mg, 8.00 mmol), and5-formyl-2,2′:5′,2″-terthiophene (221.12, 0.800 mmol) were combined withglacial acetic acid (4.0 mL) in a microwave reaction chamber and reactedwith 300 W at 180° C. for 10 minutes. The solution changed from a lightyellow colour to a deep red colour and was allowed to cool to roomtemperature. The solution was neutralized by drop-wise addition ofaqueous NH₄OH (6 mL) until the product precipitated out as ayellow/brown solid. The solid was collected using a fine glass-sinteredfrit filter and washed with H₂O (15 mL). The product was dried undervacuum to give a tan powder. (256 mg, 69%). R_(f)=(2% H₂O, 43% CHCl₃,25% Acetone, 30% MeOH+1% NH₄OH). ¹H NMR (DMSO-d₆) 7.14 (dd; 1H; J=4.37Hz), 7.33-7.43 (m; 4H), 7.55 (m; 1H), 7.75-7.79 (m; 3H), 8.82 (d; 2H;J=8.01 Hz), 8.97 (d; 2H; J=3.00 Hz).

Preparation of 14a: Ru(2,2′-bipyridine)₂Cl₂.2H₂O (156.1 mg, 0.300 mmol)and 14L (140.0 mg, 0.300 mmol) and 100% ethanol (6.0 mL) were combinedin a microwave tube and argon purged for 10 minutes by running argonthrough a needle in the cap of the tube. The cap was switched with a newone before being put in the microwave. The solution was reacted with 300W at 180° C. for 10 minutes. The resulting solution was a deep, dark redcolour. Saturated KPF₆ was added drop-wise to the aqueous solution untilno additional product precipitated (2-4 mL). The crude product wasisolated by filtration through a fine glass-sintered frit filteryielding a bright red/orange solid (307.8 mg, 88%). Purification wasdone on a silica column, eluting with a 10% H₂O:MeCN solution containing2.5% KNO₃, and the principle red spot was collected (R_(f)=0.46). Thefractions containing the desired product were combined, evaporated underreduced pressure, and further dried under vacuum to give thecorresponding NO₃ ⁻ complex and excess KNO₃. To remove the unwantedsalt, the product was dissolved in H₂O with sonication. Saturated KPF₆was added (3-4 mL), and the product precipitated out of solution. Thedesired PF6⁻ complex was extracted using CH₂Cl₂ (3×75 mL). The organiclayer was separated, concentrated under reduced pressure, and driedunder vacuum to give the final pure product as a red-orange solid (169mg, 48%). R_(f)=0.41 (10% H₂O:MeCN+2.5% KNO₃). ¹H NMR (CD₃CN): 8.65 (d;2H; J=7.89 Hz), 8.51-8.55 (m; 4H), 8.11-7.96 (m; 6H), 7.85 (d; 2H;J=5.49 Hz), 7.72 (d; 1H; J=3.81 Hz), 7.61-7.67 (m; 4H), 7.46 (t; 2H;J=6.93 Hz), 7.37 (d; 1H; J=5.10 Hz), 7.06-7.30 (m; 7H). MS (ESI+) m/z:440.0 [M-2PF₆]^(2+, 879.1) [M-2PF₆-1H]⁺. HRMS (ESI+) m/z forC₄₅H₃₀N₈RuS₃; calcd 440.0399. found 440.0382.

The PF⁶⁻ complex was dissolved in a 1:1 solution of MeCN/MeOH andconverted to its corresponding C¹⁻ salt on Amberlite IRA-410 ionexchange resin (30 cm×1.5 cm, 30 g of resin) eluting with MeOH. Anal.Calc. C₄₅H₃₀C₁₂N₈RuS₃.4.065 (H₂O): C, 52.77%; H, 3.75%; N, 10.94%.Found: C, 51.78%; H, 4.25%; N, 10.20%.

Preparation of 14C: Ru(4,4′-dimethyl-2,2′-bipyridine)₂Cl₂.2H₂O (60.2 mg,0.104 mmol) and 14L (48.5 mg, 0.104 mmol) and 100% ethanol (2.0 mL) werecombined in a microwave tube and argon purged for 15 minutes by runningargon through a needle in the cap of the tube. The cap was switched witha new one before being put in the microwave. The solution was reactedwith 300 W at 180° C. for 10 minutes. The resulting solution was a deep,dark red colour. Saturated KPF₆ was added drop-wise to the aqueoussolution until no additional product precipitated (2-3 mL). The crudeproduct was isolated by filtration through a fine glass-sintered fritfilter yielding a red/brown solid (112.6 mg, 88%). Purification was doneon a silica column, eluting with a 10% H₂O:MeCN solution containing 2.5%KNO₃, and the principle red spot was collected (R_(f)=0.54). Thefractions containing the desired product were combined, evaporated underreduced pressure, and further dried under vacuum to give thecorresponding NO₃ ⁻ complex and excess KNO₃. To remove the unwantedsalt, the product was dissolved in H₂O with sonication. Saturated KPF₆was added (3-4 mL), and the product precipitated out of solution. Thedesired PF6⁻ complex was extracted using CH₂Cl₂ (3×50 mL). The organiclayer was separated, concentrated under reduced pressure, and driedunder vacuum to give the final pure product as a red-orange solid (76.0mg, 59%). R_(f)=0.54 (10% H₂O:MeCN+2.5% KNO₃). ¹H NMR (CD₃CN): 8.87 (d;2H; J=6.60 Hz), 8.38 (d; 4H; J=10.1 Hz), 8.02 (m; 2H; J=4.50 Hz), 7.84(d; 2H; J=4.02 Hz), 7.74 (br; 2H), 7.67 (d; 2H; J=5.67 Hz), 7.53 (d; 1H;J=6.42 Hz), 7.42 (d; 2H; J=5.67 Hz), 7.25-7.31 (m; 6H), 7.07-7.14 (m;3H). MS (ESI+) m/z: 468.1 [M-2PF₆]^(2+, 1081.2) [M-PF₆]⁺. HRMS (ESI+)m/z for C₄₉H₃₈N₈RuS₃; calcd 468.0707. found 468.0697.

The PF6⁻ complex was dissolved in a 1:1 solution of MeCN/MeOH andconverted to its corresponding Cl⁻ salt on Amberlite IRA-410 ionexchange resin (30 cm×1.5 cm, 30 g of resin) eluting with MeOH.

Preparation of 16x: 2,2′-5,2″-Terthiophene-5-boronic acid pinicolester(236.1 mg, 0.780 mmol), 5-bromo-2-thiophene carboxaldyde (57.4 uL, 0.59mmol), and Pd(PPh₃)₄ (55 mg) were combined in an argon purged microwavetube. The microwave tube was again argon purged and DME (5.66 mL) and 2MNa₂CO₃ aqueous solution (0.4 mL) were then added separately by syringe.The solution was reacted at 200 W and 175° C. for 1 hour. The dark greensolution was filtered on a fine frit to remove the catalyst and washedwith tiny amounts of ethyl acetate. The filtrate was diluted withadditional EtOAc, transferred to a 125-mL reparatory, and washed withsaturated aqueous NaCl (3×50 mL). The organic layer was concentratedunder reduced pressure, and dried under vacuum to give the crude product(130.9 mg, 62%). Purification was done on a silica column, eluting with1:1 DCM:hexanes. A slow moving spot that stained positive to havealdehyde by Dinitrophenylhydrazine was collected, concentrated underreduced pressure, and dried under vacuum to give the pure product (14.1mg, 6.7%). R_(f)=0.20 (1:1 DCM:hexanes). ¹H NMR (CDCl₃) 9.86 (s; 1H),7.67 (d; 1H; J=3.96 Hz), 7.21-7.29 (m; 3H), 7.20 (d; 1H; J=3.54 Hz),7.11-7.14 (m; 3H), 7.04 (t; 1H; J=4.89 Hz).

Preparation of 16L: 1,10-Phenanthroline-5,6-dione (41.6 mg, 0.200 mmol),ammonium acetate (154 mg, 2.00 mmol), and 16× (71.7 mg, 0.200 mmol) werecombined with glacial acetic acid (1.0 mL) in a microwave reactionchamber and reacted at 300 W at 180° C. for 10 minutes. The solution wasallowed to cool to room temperature followed by neutralization bydropwise addition of aqueous NH₄OH (2-4 mL) until the productprecipitated out as a brown solid. The solid was collected using a fineglass-sintered frit filter and washed with H₂O (15 mL). The product wasdried under vacuum to give the crude product as a brown powder (105.9mg, 96%). Purification was done by recrystallization from hot MeOH togive the pure product (31.1 mg, 28%). ¹H NMR (DMSO-d₆) 9.04 (br; 2H),8.84 (d; 2H; J=7.35 Hz), 7.86 (br; 2H), 7.33-7.57 (m; 8H); 7.11 (br;1H). MS (ESI+) m/z: 549.0 [M+1H]⁺. HRMS (ESI+) m/z for C₂₉H₁₇N₄S₄; calcd549.0331. found 549.0307.

Preparation of 16a: Ru(2,2′-bipyridine)₂Cl₂.2H₂O (81.2 mg, 0.156 mmol)and 16L (85.7 mg, 0.156 mmol) and 100% ethanol (3.0 mL) were combined ina microwave tube and argon purged for 10 minutes by running argonthrough a needle in the cap of the tube. The cap was switched with a newone before being put in the microwave. The solution was reacted with 300W at 180° C. for 10 minutes. The resulting solution was a deep, dark redcolour. Saturated KPF₆ was added drop-wise to the aqueous solution untilno additional product precipitated (2-4 mL). The crude product wasisolated by filtration through a fine glass-sintered frit filteryielding a dark red/orange solid (155.2 mg, 64%). Purification was doneon a silica column, eluting with a 7% H₂O:MeCN solution containing 2.5%KNO₃, and the principle red spot was collected (R_(f)=0.46). Thefractions containing the desired product were combined, evaporated underreduced pressure, and further dried under vacuum to give thecorresponding NO₃ ⁻ complex and excess KNO₃. To remove the unwantedsalt, the product was dissolved in H₂O with sonication. Saturated KPF₆was added (3-4 mL), and the product precipitated out of solution. Thedesired PF6⁻ complex was extracted using CH₂Cl₂ (3×75 mL). The organiclayer was separated, concentrated under reduced pressure, and driedunder vacuum to give the product as a red-orange solid (169 mg, 48%).The PF6⁻ complex was dissolved in a 1:1 solution of MeCN/MeOH andconverted to its corresponding Cl⁻ salt on Amberlite IRA-410 ionexchange resin (30 cm×1.5 cm, 30 g of resin) eluting with MeOH. Theproduct was further purified on Sephadex LH-50 in methanol and theprincipal red spot was collected. The fractions containing the desiredproduct were combined, evaporated under reduced pressure, and furtherdried under vacuum to give the final product (12.7 mg, 6.5%) R_(f)=0.35(10% H₂O:MeCN+2.5% KNO₃). ¹H NMR (CD₃CN): 8.91 (d; 2H; J=7.32 Hz), 8.50(t; 4H; J=7.92 Hz), 8.08 (t; 2H; J=7.74 Hz), 7.99 (br; 4H), 7.84 (br;3H), 7.74 (br; 4H), 7.60 (br; 2H), 7.44 (t; 2H; J=5.55 Hz), 7.37 (br;1H) 7.08-7.27 (m; 9H). MS (ESI+) m/z: 498.1 [M-2PF₆]²⁺. HRMS (ESI+) m/zfor C₄₉H₃₂N₈RuS₄; calcd 481.0333. found 481.0341.

Preparation of 16c: Ru(dmb)₂Cl₂.2H₂O (88.93 mg, 0.156 mmol) and 16L(85.7 mg, 0.156 mmol) and 100% ethanol (3.0 mL) were combined in amicrowave tube and argon purged for 10 minutes by running argon througha needle in the cap of the tube. The cap was switched with a new onebefore being put in the microwave. The solution was reacted with 300 Wat 180° C. for 10 minutes. The resulting solution was a deep, dark redcolour. Saturated KPF₆ was added drop-wise to the aqueous solution untilno additional product precipitated (1-2 mL). The crude product wasisolated by filtration through a fine glass-sintered frit filteryielding a dark red (156.7 mg, 92%). Purification was done on a silicacolumn, eluting with a 7% H₂O:MeCN solution containing 2.5% KNO₃, andthe principle red spot was collected. The fractions containing thedesired product were combined, evaporated under reduced pressure, andfurther dried under vacuum to give the corresponding NO₃ ⁻ complex andexcess KNO₃. To remove the unwanted salt, the product was dissolved inH₂O with sonication. Saturated KPF₆ was added (3-4 mL), and the productprecipitated out of solution. The desired PF6⁻ complex was extractedusing CH₂Cl₂ (3×75 mL). The organic layer was separated, concentratedunder reduced pressure, and dried under vacuum to give the product as ared solid (69.9 mg, 41%). The PF6⁻ complex was dissolved in a 1:1solution of MeCN/MeOH and converted to its corresponding Cl⁻ salt onAmberlite IRA-410 ion exchange resin (30 cm×1.5 cm, 30 g of resin)eluting with MeOH. The product was further purified on Sephadex LH-50 inmethanol and the principal red spot was collected. The fractionscontaining the desired product were combined, evaporated under reducedpressure, and further dried under vacuum to give the final product (23.0mg, 13%) R_(f)=0.33 (10% H₂O:MeCN+2.5% KNO₃). ¹H NMR (CD₃CN): 8.92 (br;2H), 8.34 (s; 4H), 7.97 (br; 2H), 7.87 (br; 1H), 7.62-7.65 (m; 4H),6.92-7.42 (m; 14H), 2.54, (s; 6H), 2.46 (s; 6H). MS (ESI+) m/z: 509.1[M-2PF₆]^(2+, 1163.1) [M-PF₆]⁺. HRMS (ESI+) m/z for C₅₃H₄₀N₈RuS₄; calcd509.0646. found 509.0663.

Preparation of 2L: 1,10-Phenanthroline-5,6-dione (234.1 mg, 1.11 mmol),ammonium acetate (1.77 g, 23.0 mmol), and 2,5-thiophene dicarboxaldehyde(77.9 mg, 0.556 mmol) were combined in glacial acetic acid (20 mL) andrefluxed in air at 135° C. for 6 hours. The solution changed from alight yellow colour to a deep red colour and the product precipitated asa light orange stringy precipitate. The reaction was cooled to roomtemperature. The orange precipitate was collected using a mediumglass-sintered frit filter and washed with H₂O (10 mL). The product wasdried under vacuum to give the final pure product as a light orangepowder. No purification was needed. (90.7 mg, 31.3%). R_(f)=streak to0.60 (2% H₂O, 43% CHCl₃, 25% Acetone, 30% MeOH+1% NH₄OH). ¹H NMR (300MHz) [(CD₃)₂SO]: 9.07 (d; 4H; J=3.03 Hz), 8.90 (d; 4H; J=8.22 Hz), 8.04(s; 2H), 7.86-7.90 (br; 4H). MS (ESI+) m/z: 521.1 [M+H]⁺. HRMS (ESI+)m/z for C₃₀H₁₇N₈S; calcd 521.1291. found 521.1265.

Preparation of 3L: 1,10-Phenanthroline-5,6-dione (18.9 mg, 0.0900 mmol),ammonium acetate (138.7 mg, 1.80 mmol), and2,2′-bithiophene-5,5′-dicarbaldehyde (10 mg, 0.0450 mmol) were combinedin glacial acetic acid (5 mL) and refluxed in air at 135° C. for 6hours. The solution changed from a light yellow colour to a deep redcolour and the product precipitated as a light orange precipitatepowder. The reaction was cooled to room temperature. The solution wasneutralized by drop-wise addition of aqueous NH₄OH (5 mL) until moredesired product finished precipitating out as an orange solid. Theorange precipitate was collected using a fine glass-sintered frit filterand washed with H₂O (10 mL). The product was dried under vacuum to givean orange powder which showed the product and contaminant by NMR. (12.6mg, 47%). An alternate microwave synthesis was also performed involving1,10-Phenanthroline-5,6-dione (18.9 mg, 0.0900 mmol), ammonium acetate(138.7 mg, 1.80 mmol), and 2,2′-bithiophene-5,5′-dicarbaldehyde (10 mg,0.0450 mmol) being combined in glacial acetic acid (1.0 mL) in amicrowave reaction chamber and reacted with 300 W at 180° C. for 10minutes. The solution changed from a light yellow colour to a deep redcolour and was allowed to cool to room temperature. An orangeprecipitate of the product was seen. The solution was neutralized bydrop-wise addition of aqueous NH₄OH (1 mL) until more desired productfinished precipitating out as an orange solid. The solid was collectedusing a fine glass-sintered frit filter and washed with H₂O (10 mL). Theproduct was dried under vacuum to give an orange powder which showed theproduct and contaminant by NMR. (25.3 mg, 93%) R_(f)=streak to 0.68 (2%H₂O, 43% CHCl₃, 25% Acetone, 30% MeOH+1% NH₄OH). ¹H NMR (300 MHz)[(CD₃)₂SO]: 9.02 (br), 8.85 (br; 4H), 7.84 (m; 2H), 7.70 (br).

Preparation of 2a: Ru(bpy)₂Cl₂.2H₂O (60 mg, 0.115 mmol) and 2L (30 mg,0.0576 mmol) were combined in glycerol (3 mL) and refluxed in argon for16 hours at 100° C. The glycerol was first added to a dry reaction flaskwhich was attached to a Schlenk line. The reaction flask was evacuateduntil bubbling in the glycerol occurred and then was filled with dryargon. The evacuation and argon purging was repeated two more times.Ru(bpy)₂Cl₂.2H₂O was added to the glycerol and the reaction flask wasevacuated and argon filled three more times. The resulting glycerol andRu(bpy)₂Cl₂.2H₂O solution was heated to 100° C. for 30 minutes. Afterheating the 2L was added to the solution and the reaction flask wasevacuated and filled with argon three more times. The dark purplesolution was put under argon using a filled balloon and reacted for 16hours at 100° C. The resulting solution was a deep dark red/purplecolour and cooled to room temperature. 12 mL of H₂O was added tosolution and filtered over a medium glass-sintered frit. Saturated KPF₆was added drop-wise to the aqueous solution until no additional productprecipitated (1-2 mL). The crude product was isolated by filtrationthrough a fine glass-sintered frit filter yielding a dark red/brownsolid (106.1 mg, 0.0550 mmol). Purification was done on a silica column,eluting with a 20% H₂O:MeCN solution containing 2.5% KNO₃, and theprinciple red spot was collected. The fractions containing the desiredproduct were combined, evaporated under reduced pressure, and furtherdried under vacuum to give the product and excess KNO₃. To remove theunwanted salt, the product was dissolved in H₂O with sonication. Theproduct did not dissolve in H₂O, a quantity of H₂O (approximately 50-100mL) was added to ensure the excess KNO₃ would dissolve in the solutionfollowed by the addition of copious amounts of saturated KPF₆ (10-12mL). The desired PF6 complex was extracted using CH₂Cl₂ (3×50-150 mL).The organic layer was separated, concentrated under reduced pressure,and dried under vacuum to give the final pure product as a red-orangesolid (38.8 mg, 35%). R_(f)=0.19 (20% H₂O:MeCN+2.5% KNO3). ¹H NMR (300MHz) [(CD₃)₂SO]: 9.02-9.10 (dd; 4H; J=9.06 Hz), 8.88 (t; 8H; J=8.52 Hz),8.24 (t; 4H; J=6.60 Hz), 8.09-8.15 (m; 10H), 7.97 (br; 4H), 7.85 (d; 4H;J=4.14 Hz), 7.61 (m; 8H), 7.36 (t; 4H; J=4.68 Hz). 13C NMR [(CD3CN]158.0, 157.8, 152.8, 151.3, 148.3, 146.7, 138.7, 138.2, 135.6, 135.6,131.2, 129.0, 128.3, 126.8, 125.1, 122.1. Anal. CalcC₇₀H₄₈F₂₄N₁₆P₄Ru₂S.6 (H₂O).0.5 (C₃H₆O): C, 41.60%; H, 3.08%; N, 10.86%.Found: C, 41.74%; H, 2.72%; N, 10.16%.

The PF₆ ⁻ complex was dissolved in a 1:1 solution of MeCN/MeOH andconverted to its corresponding Cl⁻ salt on Amberlite IRA-410 ionexchange resin (30 cm×1.5 cm, 30 g of resin) eluting with MeOH. MS(ESI+) m/z: 673.1 [M-4Cl-2H]²⁺, 449.1 [M-4Cl-H]³⁺. HRMS (ESI+) m/z forC₇₀H₄₆N₁₆Ru₂S; calcd 673.0944. found 673.0945.

Preparation of 3a: Ru(bpy)₂Cl₂.2H₂O (50.0 mg, 0.0960 mmol) and 3L (30.0mg, 0.0500 mmol) were combined in glycerol (3 mL) and refluxed in argonfor 16 hours at 100° C. The glycerol was first added to a dry reactionflask which was attached to a Schlenk line. The reaction flask wasevacuated until bubbling in the glycerol occurred and then was filledwith dry argon. The evacuation and argon filling was repeated two moretimes. Ru(bpy)₂Cl₂.2H₂O was added to the glycerol and the reaction flaskwas evacuated and argon filled three more times. The resulting glyceroland Ru(bpy)₂Cl₂.2H₂O solution was heated to 100° C. for 30 minutes.After heating the glycerol the 3L was added to the solution and thereaction flask was evacuated and filled with argon three more times. Thedark purple solution was put under argon using a filled balloon andreacted for 16 hours at 100° C. The resulting solution was a deep darkred/purple colour and cooled top room temperature. 12 mL of H₂O wasadded to solution and filtered over a medium glass-sintered frit.Saturated KPF₆ was added drop wise to the filtrate until the productcompletely precipitated out of solution. The crude product was obtainedthrough filtration and washed with H₂O (5 mL). The crude product wasdark red/black in colour (78.1 mg, 0.0389 mmol). Purification was doneon a silica column, eluting with a 20% H₂O:MeCN solution containing 2.5%KNO₃, and the principle red spot was collected. The fractions containingthe desired product were combined, evaporated under reduced pressure,and further dried under vacuum to give the product and excess KNO₃. Toremove the unwanted salt, the product was attempted to be dissolved inH₂O with sonication. The product did not dissolve in H₂O and a quantityof H₂O (approximately 50 mL) was added to ensure the excess KNO₃ woulddissolve in the solution followed by the addition of copious amounts ofKPF₆ (10-12 mL). The desired PF₆ ⁻ complex was extracted using CH₂Cl₂(3×50-150 mL). The organic layer was separated, concentrated underreduced pressure, and dried under vacuum to give the final pure productas a red-orange solid (19.4 mg, 19%). R_(f)=0.27 (20% H₂O:MeCN+2.5%KNO₃). ¹H NMR (300 MHz) [(CD₃)₂SO]: 9.04 (dd; 4H; J=11.25 Hz), 8.87 (t;8H; J=10.41 Hz), 8.23 (t; 4H; J=6.57 Hz), 8.07-8.15 (m; 8H), 7.93-8.03(m; 6H), 7.84 (br; 4H), 7.74 (br; 2H), 7.60 (br; 8H), 7.36 (t; 4H;J=6.30 Hz). Anal. Calc. C₇₄H₅₀F₂₄N₁₆P₄Ru₂S₂.8(H₂O).2(C₆H₁₄): C, 44.41%;H, 4.07%; N, 9.64%. Found: C, 44.76%; H, 3.61%; N, 8.52%. MS (ESI+) m/z:714.1 [M-4PF₆.2H]²⁺, 476.4 [M-4PF₆-1H]³⁺. HRMS (ESI+) m/z forC₇₄H₄₉N₁₆Ru₂S₂; calcd 476.3946. found 476.3941. The PF6 complex wasdissolved in a 1:1 solution of MeCN/MeOH and converted to itscorresponding Cl⁻ salt on Amberlite IRA-410 ion exchange resin (30cm×1.5 cm, 30 g of resin) eluting with MeOH.

Preparation of 2b: Compound 2b was prepared by the same procedure as 2ausing Ru(phen)₂Cl₂. ¹H NMR (300 MHz) [(CD₃CN]: 9.01 (d; 2H; J=10.17 Hz),8.86 (d; 2H; J=7.68 Hz), 8.63 (d; 8H; J=7.98 Hz), 8.29 (s; 8H), 8.13(br; 4H), 8.00-8.04 (m; 10H), 7.66 (m; 12H). Anal. Calc.C₃₇H₂₆F₁₂N₈P₂RuS.0.5 (H₂O): C, 43.79%; H, 2.68%; N, 11.04%. Found: C,44.66%; H, 2.98%; N, 10.16%. The PF₆ ⁻ complex was dissolved in a 1:1solution of MeCN/MeOH and converted to its corresponding Cl⁻ salt onAmberlite IRA-410 ion exchange resin (30 cm×1.5 cm, 30 g of resin)eluting MeOH. MS (ESI+) m/z: 721.1 [M-4Cl-2H]²⁺, 481.1 [M-4Cl-H]³⁺. HRMS(ESI+) m/z for C₇₈H₄₆N₁₆Ru₂S; calcd 721.0944. found 721.0971.

Preparation of 3b: Compound 3b was prepared by the same procedure as 2ausing Ru(phen)₂Cl₂. ¹H NMR (300 MHz) [CD₃CN]: 8.94 (d; 4H; J=20.31 Hz),8.63 (d; 8H; J=7.95 Hz), 8.28 (s; 8H), 8.11 (br; 4H), 8.04 (br; 4H),7.99 (br; 4H), 7.93 (br; 2H), 7.63-7.70 (m; 12H), 7.56 (br; 2H). Anal.Calc. C₈₂H₅₀F₂₄N₁₆P₄Ru₂S₂.3(C₆H₁₄).8(CH₃OH): C, 49.50%; H, 4.77%; N,8.55%. Found: C, 49.15%; H, 3.94%; N, 7.64%. MS (ESI+) m/z: 908.1[M-2PF₆]²⁺, 508.4 [M-4 PF₆-1H]³⁺, 381.5 [M-4 PF₆]⁴⁺. HRMS (ESI+) m/z forC₈₂H₅₀F₁₂N₁₆P₂Ru₂S₂; calcd 908.0603. found 908.0641. The PF₆ ⁻ complexwas dissolved in a 1:1 solution of MeCN/MeOH and converted to itscorresponding Cl⁻ salt on Amberlite IRA-410 ion exchange resin (30cm×1.5 cm, 30 g of resin) eluting MeOH.

Preparation of 10b: Compound 10b was prepared by the same procedure as2a using Ru(phen)₂Cl₂. ¹H NMR (DMSO-d₆) δ 7.17 (m, 1H), 7.50 (m, 2H),7.63 (d, 1H), 7.77 (m, 6H), 7.90 (m, 1H), 7.99 (d, 2H), 8.07 (d, 2H),8.12 (d, 2H), 8.40 (s, 4H), 8.78 (d, 4H), 8.99 (d, 2H). MS (ESI+) m/z:845.0 [M-2PF₆]⁺, 423.0 [M-2PF₆]²⁺. HRMS (ESI+) m/z for C₄₅H₂₈N₈RuS₂;calcd 846.0922. found 846.0910. The PF₆ ⁻ complex was dissolved in a 1:1solution of MeCN/MeOH and converted to its corresponding Cl⁻ salt onAmberlite IRA-410 ion exchange resin (30 cm×1.5 cm, 30 g of resin)eluting MeOH.

Preparation of 14b: Compound 14b was prepared by the same procedure as2a using Ru(phen)₂Cl₂. ¹H NMR (300 MHz) [CD₃CN]: 8.56-8.63 (m; 6H; C, 7,4), 8.31-8.26 (m; 8H; 6, 5, 9), 8.01 (d; 2H; J=5.10 Hz; 2), 7.88 (d; 2H;J=4.95 Hz; A), 7.72 (br; 2H; B), 7.60 (m; 3H; 8, 1H—R), 7.29-7.40 (m;3H; 3, 1H—R), 7.02 (m; 2H; 2H—R), 6.87 (d; 1H; J=3.72 Hz; 1H—R),6.68-7.73 (m; 2H; 2H—R). MS (ESI+) m/z: 464.0 [M-2PF₆]²⁺. HRMS (ESI+)m/z for C₄₉H₃₀N₈RuS₃; calcd 464.0400. found 464.0381. The PF₆ ⁻ complexwas dissolved in a 1:1 solution of MeCN/MeOH and converted to itscorresponding Cl⁻ salt on Amberlite IRA-410 ion exchange resin (30cm×1.5 cm, 30 g of resin) eluting MeOH. Anal. Calc.C₄₉H₃₀N₈Cl₂RuS₃.4(H₂O).6(CH₃OH): C, 52.29%; H, 4.95%; N, 8.87%. Found:C, 53.11%; H, 4.31%; N, 8.33%.

FORMULATIONS

The present invention also relates to compositions or formulations whichcomprise the photodynamic compounds according to the present invention.In general, the compositions of the present invention comprise aneffective amount of one or more photodynamic compounds and salts thereofaccording to the present invention which are effective for providingphotodynamic therapy; and one or more excipients.

For the purposes of the present invention the term “excipient” and“carrier” are used interchangeably throughout the description of thepresent invention and said terms are defined herein as, “ingredientswhich are used in the practice of formulating a safe and effectivepharmaceutical composition.”

The formulator will understand that excipients are used primarily toserve in delivering a safe, stable, and functional pharmaceutical,serving not only as part of the overall vehicle for delivery but also asa means for achieving effective absorption by the recipient of theactive ingredient. An excipient may fill a role as simple and direct asbeing an inert filler, or an excipient as used herein may be part of apH stabilizing system or coating to insure delivery of the ingredientssafely to the stomach. The formulator can also take advantage of thefact the compounds of the present invention have improved cellularpotency, pharmacokinetic properties, as well as improved oralbioavailability.

The present teachings also provide pharmaceutical compositions thatinclude at least one compound described herein and one or morepharmaceutically acceptable carriers, excipients, or diluents. Examplesof such carriers are well known to those skilled in the art and can beprepared in accordance with acceptable pharmaceutical procedures, suchas, for example, those described in Remington's Pharmaceutical Sciences,17th edition, ed. Alfonoso R. Gennaro, Mack Publishing Company, Easton,Pa. (1985), the entire disclosure of which is incorporated by referenceherein for all purposes. As used herein, “pharmaceutically acceptable”refers to a substance that is acceptable for use in pharmaceuticalapplications from a toxicological perspective and does not adverselyinteract with the active ingredient. Accordingly, pharmaceuticallyacceptable carriers are those that are compatible with the otheringredients in the formulation and are biologically acceptable.Supplementary active ingredients can also be incorporated into thepharmaceutical compositions.

Compounds of the present teachings can be administered orally orparenterally, neat or in combination with conventional pharmaceuticalcarriers. Applicable solid carriers can include one or more substanceswhich can also act as flavoring agents, lubricants, solubilizers,suspending agents, fillers, glidants, compression aids, binders ortablet-disintegrating agents, or encapsulating materials. The compoundscan be formulated in conventional manner, for example, in a mannersimilar to that used for known photodynamic compounds. Oral formulationscontaining a compound disclosed herein can comprise any conventionallyused oral form, including tablets, capsules, buccal forms, troches,lozenges and oral liquids, suspensions or solutions. In powders, thecarrier can be a finely divided solid, which is an admixture with afinely divided compound. In tablets, a compound disclosed herein can bemixed with a carrier having the necessary compression properties insuitable proportions and compacted in the shape and size desired. Thepowders and tablets can contain up to 99% of the compound.

Capsules can contain mixtures of one or more compound(s) disclosedherein with inert filler(s) and/or diluent(s) such as pharmaceuticallyacceptable starches (e.g., corn, potato or tapioca starch), sugars,artificial sweetening agents, powdered celluloses (e.g., crystalline andmicrocrystalline celluloses), flours, gelatins, gums, and the like.

Useful tablet formulations can be made by conventional compression, wetgranulation or dry granulation methods and utilize pharmaceuticallyacceptable diluents, binding agents, lubricants, disintegrants, surfacemodifying agents (including surfactants), suspending or stabilizingagents, including, but not limited to, magnesium stearate, stearic acid,sodium lauryl sulfate, talc, sugars, lactose, dextrin, starch, gelatin,cellulose, methyl cellulose, microcrystalline cellulose, sodiumcarboxymethyl cellulose, carboxymethylcellulose calcium,polyvinylpyrrolidine, alginic acid, acacia gum, xanthan gum, sodiumcitrate, complex silicates, calcium carbonate, glycine, sucrose,sorbitol, dicalcium phosphate, calcium sulfate, lactose, kaolin,mannitol, sodium chloride, low melting waxes, and ion exchange resins.Surface modifying agents include nonionic and anionic surface modifyingagents. Representative examples of surface modifying agents include, butare not limited to, poloxamer 188, benzalkonium chloride, calciumstearate, cetostearl alcohol, cetomacrogol emulsifying wax, sorbitanesters, colloidal silicon dioxide, phosphates, sodium dodecylsulfate,magnesium aluminum silicate, and triethanolamine. Oral formulationsherein can utilize standard delay or time-release formulations to alterthe absorption of the compound(s). The oral formulation can also consistof administering a compound disclosed herein in water or fruit juice,containing appropriate solubilizers or emulsifiers as needed.

Liquid carriers can be used in preparing solutions, suspensions,emulsions, syrups, elixirs, and for inhaled delivery. A compound of thepresent teachings can be dissolved or suspended in a pharmaceuticallyacceptable liquid carrier such as water, an organic solvent, or amixture of both, or a pharmaceutically acceptable oils or fats. Theliquid carrier can contain other suitable pharmaceutical additives suchas solubilizers, emulsifiers, buffers, preservatives, sweeteners,flavoring agents, suspending agents, thickening agents, colors,viscosity regulators, stabilizers, and osmo-regulators. Examples ofliquid carriers for oral and parenteral administration include, but arenot limited to, water (particularly containing additives as describedherein, e.g., cellulose derivatives such as a sodium carboxymethylcellulose solution), alcohols (including monohydric alcohols andpolyhydric alcohols, e.g., glycols) and their derivatives, and oils(e.g., fractionated coconut oil and arachis oil). For parenteraladministration, the carrier can be an oily ester such as ethyl oleateand isopropyl myristate. Sterile liquid carriers are used in sterileliquid form compositions for parenteral administration. The liquidcarrier for pressurized compositions can be halogenated hydrocarbon orother pharmaceutically acceptable propellants.

Liquid pharmaceutical compositions, which are sterile solutions orsuspensions, can be utilized by, for example, intramuscular,intraperitoneal or subcutaneous injection. Sterile solutions can also beadministered intravenously. Compositions for oral administration can bein either liquid or solid form.

Preferably the pharmaceutical composition is in unit dosage form, forexample, as tablets, capsules, powders, solutions, suspensions,emulsions, granules, or suppositories. In such form, the pharmaceuticalcomposition can be sub-divided in unit dose(s) containing appropriatequantities of the compound. The unit dosage forms can be packagedcompositions, for example, packeted powders, vials, ampoules, prefilledsyringes or sachets containing liquids. Alternatively, the unit dosageform can be a capsule or tablet itself, or it can be the appropriatenumber of any such compositions in package form. Such unit dosage formcan contain from about 1 mg/kg of compound to about 500 mg/kg ofcompound, and can be given in a single dose or in two or more doses.Such doses can be administered in any manner useful in directing thecompound(s) to the recipient's bloodstream, including orally, viaimplants, parenterally (including intravenous, intraperitoneal andsubcutaneous injections), rectally, vaginally, and transdermally.

When administered for the treatment or inhibition of a particulardisease state or disorder, it is understood that an effective dosage canvary depending upon the particular compound utilized, the mode ofadministration, and severity of the condition being treated, as well asthe various physical factors related to the individual being treated. Intherapeutic applications, a compound of the present teachings can beprovided to a patient already suffering from a disease in an amountsufficient to cure or at least partially ameliorate the symptoms of thedisease and its complications. The dosage to be used in the treatment ofa specific individual typically must be subjectively determined by theattending physician. The variables involved include the specificcondition and its state as well as the size, age and response pattern ofthe patient.

In some cases it may be desirable to administer a compound directly tothe airways of the patient, using devices such as, but not limited to,metered dose inhalers, breath-operated inhalers, multidose dry-powderinhalers, pumps, squeeze-actuated nebulized spray dispensers, aerosoldispensers, and aerosol nebulizers. For administration by intranasal orintrabronchial inhalation, the compounds of the present teachings can beformulated into a liquid composition, a solid composition, or an aerosolcomposition. The liquid composition can include, by way of illustration,one or more compounds of the present teachings dissolved, partiallydissolved, or suspended in one or more pharmaceutically acceptablesolvents and can be administered by, for example, a pump or asqueeze-actuated nebulized spray dispenser. The solvents can be, forexample, isotonic saline or bacteriostatic water. The solid compositioncan be, by way of illustration, a powder preparation including one ormore compounds of the present teachings intermixed with lactose or otherinert powders that are acceptable for intrabronchial use, and can beadministered by, for example, an aerosol dispenser or a device thatbreaks or punctures a capsule encasing the solid composition anddelivers the solid composition for inhalation. The aerosol compositioncan include, by way of illustration, one or more compounds of thepresent teachings, propellants, surfactants, and co-solvents, and can beadministered by, for example, a metered device. The propellants can be achlorofluorocarbon (CFC), a hydrofluoroalkane (HFA), or otherpropellants that are physiologically and environmentally acceptable.

Compounds described herein can be administered parenterally orintraperitoneally. Solutions or suspensions of these compounds or apharmaceutically acceptable salts, hydrates, or esters thereof can beprepared in water suitably mixed with a surfactant such ashydroxyl-propylcellulose. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, and mixtures thereof in oils. Underordinary conditions of storage and use, these preparations typicallycontain a preservative to inhibit the growth of microorganisms.

The pharmaceutical forms suitable for injection can include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In certain embodiments, the form can sterile and itsviscosity permits it to flow through a syringe. The form preferably isstable under the conditions of manufacture and storage and can bepreserved against the contaminating action of microorganisms such asbacteria and fungi. The carrier can be a solvent or dispersion mediumcontaining, for example, water, ethanol, polyol (e.g., glycerol,propylene glycol and liquid polyethylene glycol), suitable mixturesthereof, and vegetable oils.

Compounds described herein can be administered transdermally, i.e.,administered across the surface of the body and the inner linings ofbodily passages including epithelial and mucosal tissues. Suchadministration can be carried out using the compounds of the presentteachings including pharmaceutically acceptable salts, hydrates, oresters thereof, in lotions, creams, foams, patches, suspensions,solutions, and suppositories (rectal and vaginal).

Transdermal administration can be accomplished through the use of atransdermal patch containing a compound, such as a compound disclosedherein, and a carrier that can be inert to the compound, can benon-toxic to the skin, and can allow delivery of the compound forsystemic absorption into the blood stream via the skin. The carrier cantake any number of forms such as creams and ointments, pastes, gels, andocclusive devices. The creams and ointments can be viscous liquid orsemisolid emulsions of either the oil-in-water or water-in-oil type.Pastes comprised of absorptive powders dispersed in petroleum orhydrophilic petroleum containing the compound can also be suitable. Avariety of occlusive devices can be used to release the compound intothe blood stream, such as a semi-permeable membrane covering a reservoircontaining the compound with or without a carrier, or a matrixcontaining the compound. Other occlusive devices are known in theliterature.

Compounds described herein can be administered rectally or vaginally inthe form of a conventional suppository. Suppository formulations can bemade from traditional materials, including cocoa butter, with or withoutthe addition of waxes to alter the suppository's melting point, andglycerin. Water-soluble suppository bases, such as polyethylene glycolsof various molecular weights, can also be used.

Lipid formulations or nanocapsules can be used to introduce compounds ofthe present teachings into host cells either in vitro or in vivo. Lipidformulations and nanocapsules can be prepared by methods known in theart.

To increase the effectiveness of compounds of the present teachings, itcan be desirable to combine a compound with other agents effective inthe treatment of the target disease. For example, other active compounds(i.e., other active ingredients or agents) effective in treating thetarget disease can be administered with compounds of the presentteachings. The other agents can be administered at the same time or atdifferent times than the compounds disclosed herein.

Compounds of the present teachings can be useful for the treatment orinhibition of a pathological condition or disorder in a mammal, forexample, a human subject. The present teachings accordingly providemethods of treating or inhibiting a pathological condition or disorderby providing to a mammal a compound of the present teachings includingits pharmaceutically acceptable salt) or a pharmaceutical compositionthat includes one or more compounds of the present teachings incombination or association with pharmaceutically acceptable carriers.Compounds of the present teachings can be administered alone or incombination with other therapeutically effective compounds or therapiesfor the treatment or inhibition of the pathological condition ordisorder.

Non-limiting examples of compositions according to the present inventioninclude from about 0.001 mg to about 1000 mg of one or more photodynamiccompounds according to the present invention and one or more excipients;from about 0.01 mg to about 100 mg of one or more photodynamic compoundsaccording to the present invention and one or more excipients; and fromabout 0.1 mg to about 10 mg of one or more photodynamic compoundsaccording to the present invention; and one or more excipients.

Procedures

The following procedures can be utilized in evaluating and selectingcompounds as photodynamic compounds.

Extraction of topoisomerase II activity from HL60 cells

Nuclear extracts were affinity precipitated as described in ‘Small ScalePreparation of Topo I and II Extracts from Tissue Culture Cells(Optimized for HeLa Cells)’ on the TopoGEN website(http:www.topogen.com/html/extracts.html). All steps were conducted onice or at 4° C. Briefly, 10 mL of exponentially growing HL-60 cells(1×10⁶ cells/mL) were transferred to a sterile 15 mL conical centrifugetube (Fisher Scientific, Canada) and pelleted in an eppendorf 5804Rcentrifuge (16.1 cm radius) at 2100 rpm for 3 min at 4° C. The cellswere washed twice with 3 mL of ice cold phosphate buffered saline (PBS)containing 2.68 mM potassium chloride, 1.47 mM potassium phosphatemonobasic, 0.137 M sodium chloride, and 8.10 mM sodium phosphatedibasic, pH 7.4, as follows: the cell pellet was resuspended with PBSbuffer (mixed by pipetting up and down), centrifuged at 2100 rpm (3 min,4° C.), and the supernatant gently poured off. After the second wash,the cells were resuspended in 3 mL of cold hypotonic buffer (10 mMTris-HCl, pH 7.5, 1 mM EDTA, 4 mM MgCl₂, 0.5 mM phenylmethylsulfonylfluoride (PMSF)), and clumps were dispersed by pipetting up and down.The cells were pelleted again (2100 rpm, 3 min, 4° C.), resuspended anddispersed in 3 mL of the same cold hypotonic buffer, and left on ice for10 min to swell. Cell membranes were disrupted with a cold Douncehomogenizer (Pyrex, 15 mL), using 6-8 strokes. The lysate wastransferred to a clean sterile 1.5 mL microcentrifuge tube (FisherScientific, Canada) and centrifuged (2900 rpm, 10 min, 4° C.). Thepellet, containing nuclei, was washed twice with the same cold hypotonicbuffer, as follows: cold buffer was added to the pellet and the nucleiwas resuspended by pipetting up and down, then pelleted (2900 rpm, 10min, 4° C.), the supernatant gently removed and discarded. After thesecond wash, the nuclei were resuspended in 4 pellet volumes(approximately 500 μL) of cold hypotonic buffer without MgCl₂. An equalvolume of cold 1 M NaCl was added to the resuspended pellet and left onice for 45 min, followed by pelleting in a microcentrifuge (14,000 rpm,15 min, 4° C.). The supernatant (nuclear extract), suspended in 5 mMTris-HCl (pH 7.5), 0.5 mM EDTA, 0.25 mM PMSF, and 0.5 M NaCl, was usedfor DNA relaxation assays.

Total protein concentration (BSA equivalent) of the extract wasdetermined by using a Micro Lowry total protein kit (Total Protein Kit,Micro Lowry, Peterson's modification), following the manufacturer'sinstruction for ‘Protein Determination without Protein Precipitation.’Briefly, five BSA protein standards were prepared from a 400 μg/mL stocksolution in pure water to a volume of 200 μL in 1.5 mL sterilemicrocentrifuge tubes (final concentrations of BSA were 10, 20, 40, 60,80 μg). A sixth tube containing pure water only was used as a blank. Aseventh tube, containing 50 μL nuclear extract and 150 μL (dilutionchosen randomly) was prepared in order to measure its protein contentagainst the BSA standards. All tubes were mixed well with vortexing,then 200 μL Lowry reagent solution was added to all tubes followed byvortexing to mix. The solutions sat at room temperature for 20 min, then100 μL Folin Ciocalteu phenol reagent (6× dilution of 2 N stocksolution) was added to each tube, followed by vortexing to mix. Thecolour was allowed to develop for 30 min. The solutions were transferredone at a time to a quartz cuvette, with a pathlength of 1 cm, and theabsorbance of the standards and sample tubes versus the blank weremeasured at a wavelength of 750 nm. A plot was constructed (Excel, 2007)of the absorbance values of the standards versus their correspondingprotein concentrations and linear regression was used to calculate theprotein concentration in the nuclear extract sample, taking into accountthe 10× dilution. The result was a protein concentration of 559 μg/mL or279 μg (BSA equivalents) of the nuclear extract.

Topoisomerase Extract Activity in DNA Relaxation Assays:

Relaxation activity of the nuclear extract, containing topo I and II,was determined by detecting the conversion of supercoiled plasmid DNA toits relaxed form in the presence of ATP. Reaction tubes (20 μL volumes)were assembled on ice by the ordered addition of: (i) pure water(variable, made up to 20 μL volume); (ii) 4 μL relaxation buffer (250 mMTris-HCl, pH 8, 0.75 M NaCl, 50 mM MgCl₂, 2.5 mM dithiothreitol, 150 μgBSA/mL, and 10 mM ATP); (iii) pUC19 supercoiled plasmid DNA (250 ng, or38.6 μM bases); and (iv) topo extract (1, 2, 3, or 4 μL). The reactionwas initiated by heating the tubes in a 37° C. incubator for 30 min. Thereaction was stopped by adding 2 μL of 10% SDS (in sterile water), andthe DNA-bound protein was then digested by adding 2 μL proteinase K(0.50 mg/mL stock in 10 mM Tris-HCl, 1 mM EDTA, pH 7.5) and incubatingat 37° C. for 15 min. Then, 2 μL loading dye with ficoll (0.25%bromophenol blue, 15% ficoll, in 1×TBE buffer) was added and DNA sampleswere then analyzed by 1.5% agarose gel electrophoresis using 1×TBEbuffer (50 V, 180 min). The gels were stained with ethidium bromide (1μg/mL) for 30 min with subsequent destaining for 30 min in water, andvisualized with UV-transillumination (UVP transilluminator) using theGel-Doc-It Imaging system (UVP). One unit of DNA topoisomerase IIactivity was defined as the amount of enzyme capable of relaxing 250 ngof supercoiled DNA in 30 minutes at 37° C. (in this case, one unit=2 μLextract). The presence of topo I was assessed by testing for relaxationof pUC19 plasmid (250 ng) in the absence of ATP for 30 min at 37° C.Under these conditions, relaxation of pUC19 plasmid was not detected(indicating little to no topo I).

Topoisomerase II Assays:

Inhibition of topoisomerase II activity by compounds of the disclosurewas measured by a supercoiled DNA relaxation assay using a topoisomeraseII drug screening kit (TopoGEN). Briefly, 0.23 μg supercoiled pUC19plasmid DNA (3.5 μL of 64.5 ng/μL stock solution in 10 mM Tris-C1, pH8.5) was suspended in pH 8.0 reaction buffer (250 mM Tris-HCl, 0.75 MNaCl, 50 mM MgCl₂, 2.5 mM dithiothreitol, 150 μg BSA/mL, and 10 mM ATP).Pure water was added (variable, made up to 20 μL volume), then 2 μLaliquots of ruthenium compounds (1, 10, 50, 100, 500, 1000 μM serialdilutions) were added, making final sample concentrations of 0.1, 1, 5,10, 50, and 100 μM. Control samples were prepared as follows: (i)plasmid only (no nuclear extract); (ii) plasmid with nuclear extract;(iii) plasmid with the highest ruthenium concentration (no nuclearextract); and (iv) plasmid with nuclear extract (no ATP in the buffer).The tubes were mixed well (gently shaken and spun down) prior toinitiating the reaction by the addition of 2 μL (one unit) nuclearextract. After 30 min incubation at 37° C., the reaction was stopped byadding 2 μL of 10% SDS (in sterile water). The DNA-bound protein wasdigested by adding 2 μL of proteinase K (0.50 mg/mL stock in 10 mMTris-HCl, 1 mM EDTA, pH 7.5) at 37° C. for 15 min, without the optionalchloroform:isoamyl alcohol extraction (earlier extractions showed nocosmetic improvement in results). Lastly, 2 μL ficoll loading dye (0.25%bromophenol blue, 15% ficoll, in 1×TBE buffer) was added and DNA sampleswere analyzed by 1.5% agarose gel electrophoresis using 1×TBE buffer (50V, 180 min). The gel was stained with ethidium bromide (1 μg/mL) for 30min with subsequent destaining for 30 min in pure water, and visualizedwith UV-transillumination (UVP transilluminator) using the Gel-Doc-ItImaging system (UVP).

DNA Binding by UV-Vis:

Optical titrations were carried out on 0.5-2 mL solutions of the PDCswith increasing amounts of calf thymus or herring sperm DNA to give [DNAbases]/[PDC] between 0.1 and 10. DNA was added in 1-5 μL increments tosolutions of compound (10 μM) in 10 mM MOPS, 10 mM MOPS with 50 mM NaCl,5 mM Tris-HCl, or 5 mM Tris-HCl with 50 mM NaCl at pH 7.5. The dilutionof compounds of the disclosure at the end of each titration, althoughnegligible, was accounted for in the binding constant analyses. The DNAbinding constant (K_(b)) was obtained from fits of the titration data toeq. 1 (FIG. 36), where b=1+K_(b)C_(t)+K_(b)[DNA]_(t)/2_(s), C_(t) and[DNA]_(t) represent the total PDC and DNA concentrations, respectively,s is the binding site size, and ε_(a), ε_(f), and ε_(b) represent themolar extinction coefficients of the apparent, free, and bound PDCs,respectively. ε_(f) was calculated at 414 nm for 7 and 412 nm for 8before the addition of DNA, and ε_(a) was determined at thesewavelengths after each addition of DNA. The value of ε_(b), wasdetermined from the plateau of the DNA titration, where addition of DNAdid not result in any further decrease in the absorption signal.Detailed fits of the titration data were obtained using bothKaleidagraph and Gnuplot.

DNA melting curves were constructed by measuring the absorbance (A₂₆₀)of a 2 mL, 25 μM DNA solution (40 mM MOPS, pH 7.5) as a function oftemperature (20-100° C.) in the absence and presence of a compound ofthe disclosure (5 μM). Solutions of DNA and a compound of the disclosurefor melting experiments were allowed to equilibrate for 30 min at 25° C.prior to measurement. The ETC-505T temperature controller was cooledwith ice water (4° C.) using a fish-aquarium pump, and a stream of argongas was supplied via the gas inlet valve to the sample compartment toprevent condensation on the cuvette windows during variable-temperatureexperiments.

Photocleavage Titrations:

DNA photocleavage experiments were performed according to a generalplasmid DNA assay with 20 μL total sample volumes in 0.5 or 1.5 mLmicrofuge tubes containing transformed pUC19 plasmid (200 ng, >95% FormI) in 10 mM MOPS buffer and 100 mM NaCl, pH 7.4. DNA (1-5 μL) wasdelivered to the assay tubes as a solution in 10 mM Tris-Cl (pH 8.5) anddiluted with MOPS (pH 7.5, final concentration 10 mM) and NaCl (finalconcentration 100 mM). Solutions of the compounds of the disclosure wereadded to give the from 0 to 500 μM, and the reaction mixtures werediluted to a final volume of 20 μL, when necessary, with ddH₂O.Complexes were dissolved initially in acetonitrile (2 μM stocksolutions), and all subsequent dilutions were made with ddH₂O wherefinal assay tubes contained <1% acetonitrile. For concentration-basedassays, samples (no pre-incubation period) were irradiated in air for 30min with 420 nm light inside a photoreactor (Luzchem LZC-4×). Whereirradiation of deoxygenated samples was required, argon was bubbledthrough the solutions for 15 min prior to irradiation under a positivepressure of argon. All samples were quenched by the addition of gelloading buffer (4 μL), loaded onto 1% agarose gels containing ethidiumbromide (0.75 μg mL⁻¹), and electrophoresed for 30 min at 8-12 V cm⁻¹ in1×TAE (40 mM Trisacetate, 1 mM EDTA, pH 8.2). The bands were visualizedwith UV-transillumination (UVP transilluminator) and quantified usingthe Get Doc-It Imaging system (UVP) or GNU Image Manipulation Program(GIMP).

HL-60 Cell Culture:

HL-60 human promyelocytic leukemia cells (ATCC CCL-240) were cultured at37° C. under 5% CO₂ in Hyclone's IMDM, supplemented with 20% FBS andwere passaged 3-4 times per week according to standard asepticprocedures. Cultures were started at 200,000 cells mL⁻¹ in 25 cm² tissueculture flasks and were subcultured before growth reached 750,000 cellsmL⁻¹ to avoid senescence associated with prolonged high cell density.Complete media was prepared in 200 mL portions as needed by combiningIMDM (160 mL), FBS (40 mL, pre-aliquoted and heat inactivated), andgentamicin sulfate (100 μL of 50 mg mL⁻¹ stock solution) in a 250 mLMillipore vacuum stericup (0.22 μm) and filtering.

Cytotoxicity and Photocytotoxicity Assays

HL-60 cells growing in log phase (approximately 8×10⁵) were transferredin 50 μL aliquots to two 96-well tissue-culture microplates (CorningCostar, Acton, Mass.) containing 100 μL warm culture medium and placedin a 37° C., 5% CO₂ water-jacketed incubator (Thermo Electron Corp.,Form a Series II, Model 3110, HEPA Class 100) for one hour toequilibrate. All empty microplate wells contained 200 μL phosphatebuffered saline (PBS) containing 2.68 mM potassium chloride, 1.47 mMpotassium phosphate monobasic, 0.137 M sodium chloride, and 8.10 mMsodium phosphate dibasic, pH 7.4, to help minimize evaporation loss.Warm 50 μL aliquots of solution of compounds of the disclosure (4, 20,40, 200 μM), freshly made in PBS, were added to the cells and incubatedat 37° C. under 5% CO₂ for 4 hr (final concentrations were 1, 5, 10, 50μM). One of the microplates was irradiated with visible light (400-700nm) in a Luzchem photoreactor (cool white fluorescent tubes, 21 W/m²)for 15 min; the other microplate was incubated under identicalconditions in the dark. Both microplates were then incubated (37° C.under 5% CO₂) for 40 hr. A Cellometer Auto T4 (ESBE Scientific) was usedto determine the cell number, viability, diameter, and % cell viability.Cell suspensions (20 μL) were diluted 1:1 with 0.2% trypan blue dye(Sigma Aldrich, Canada), loaded into a cell counting chamber-slide, andinserted into the imaging-based automatic cell counter. Cellconcentration and cell viability were automatically determined(Cellometer Auto Counter Software) on the basis of the total cell count,the dilution factor, and the trypan blue dye exclusion. The optimal celltype parameters were established by importing the settings for HL-60cells, and the data was subsequently imported into a Microsoft excelspreadsheet (Microsoft Office 2010) for data analysis.

Cytotoxicity and Photocytotoxicity Assays:

HL-60 cells growing in log phase were transferred (typically 25 μLaliquots) to 96-well tissue-culture plates (Corning Costar, Acton,Mass.) containing culture medium either with or without varyingconcentrations of the compounds of the disclosure to give final volumesin the culture wells of 100 μL and 10,000-20,000 cells. Solutions ofcompounds of the disclosure in complete media were prepared in 1 mLportions, where acetonitrile from the initial compounds of thedisclosure stock solution was <0.5% v/v, and sterile-filtered in 3 mLsyringes equipped with 0.22 μm Nalgene filters. Plates were incubated at37° C. under 5% CO₂ for 30 min prior to exposure to 420 nm light in aLuzchem photoreactor for 30 min; dark controls were incubated underidentical conditions in the dark. Dark controls, or cytotoxicity (CT)assays, refer to assays that include compounds of the disclosure butwere not exposed to light, and light controls refer to light-exposedassays that did not contain compounds of the disclosure.Photocytotoxicity (PCT) assays contained PDC and were exposed to light.Cell counts and viability staining were carried out immediately and at˜24 h following light exposure. Manual counts were performed on 25 μL1:1 mixtures of assay culture and trypan Hue solution in a Neubauerhemocytometer viewed under an inverted light microscope inphase-contrast mode (4× objective, 60× total magnification). Under theseconditions, viable cells appeared bright white, and non-viable cellswere blue. All experiments were carried out in triplicate, and thegraphed data is the average of three trials.

Viability Staining:

Viability was established according to a published protocol whereby a100× stock solution of ethidium bromide/acridine orange (EB/AO) wasprepared by dissolving ethidium bromide (50 mg) and acridine orange (15mg) in 95% ethanol (1 mL) and diluting 1/50 with ddH₂O. The 100×solution was divided into 1 mL aliquots and stored at −20° C. A 1×working solution was made by thawing a 1 mL aliquot of the 100× stocksolution and diluting 1/100 with phosphate-buffered saline. The workingsolution was stored in an amber bottle at 4° C. for up to 1 month. Forcellular viability staining, an aliquot of cell suspension was adjustedto 1-5×10⁶ cells mL⁻ in phosphate-buffered IMDM. A 25 aliquot of thiscell suspension was mixed with 1×EB/AO staining solution (25 in amicrofuge tube; a 25 μL aliquot of this cell-stain mixture wastransferred to a hemocytometer and viewed under a Nikon Eclipse TE2000-Uinverted light microscope operating in epi-fluorescence mode (10× or 40×objective, 150× or 600× total magnification). Under these conditions,viable cells took up AO and excluded EB, resulting in only greenfluorescence with UV-excitation, Nonviable cells (apoptotic or necrotic)assimilated EB and fluoresced red with green excitation, overwhelmingany green fluorescence from AO. Apoptotic cells were discerned from theformation of smaller, apoptotic bodies that fluoresced red.

Nuclear Staining for Laser Scanning Confocal Microscopy (LSCM)

HL-60 human promyelocytic leukemia cells (ATCC CCL-240) growing in logphase were transferred in aliquots of 100 μL (approximately 50,000cells) to a 96-well tissue-culture microplate (Corning Costar, Acton,Mass.) containing 150 μL warm culture medium (Hyclone's IMDMsupplemented with 20% FBS), and placed in a 37° C., 5% CO₂water-jacketed incubator (Thermo Electron Corp., Form a Series II, Model3110, HEPA Class 100) for one hour to equilibrate. Then, 50 μL of a 600μM solution of a compounds of the disclosure (warmed to 37° C.), made inphosphate buffered saline (PBS) containing 2.68 mM potassium chloride,1.47 mM potassium phosphate monobasic, 0.137 M sodium chloride, and 8.10mM sodium phosphate dibasic, pH 7.4, was added. The microplate wasreturned to the incubator for 15 min. The 300 μL sample was thentransferred to a collagen-coated glass-bottom tissue culture dish(FluoroDish FD35COL, World Precision Instruments Inc.) and returned tothe incubator for 10-15 min to allow cells to adhere to the coated dish.The volume of the tissue culture dish was subsequently topped up to 2 mLwith warm PBS for LSCM.

While the invention has been described in detail and with reference tospecific examples thereof, it will be apparent to one skilled in the artthat various changes and modifications can be made therein withoutdeparting from the spirit and scope thereof.

What is claimed is:
 1. A compound having formula (I):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein: M is selected from the groupconsisting of manganese, molybdenum, rhenium, iron, ruthenium, osmium,cobalt, rhodium, iridium, nickel, platinum, and copper; X is selectedfrom the group consisting of Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄ ⁻, CF₃SO₃ ⁻,and SO₄ ⁻²; n=0, 1, 2, 3, 4, or 5; y=1, 2, or 3; z=0, 1, or 2; Lig ateach occurrence is independently selected from the group consisting of

R¹ is selected from the group consisting of

u is an integer; R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), and R^(2f) ateach occurrence are each independently selected from the groupconsisting of hydrogen, C1-6 optionally substituted alkyl, C1-6optionally substituted branched alkyl, C3-7 optionally substitutedcycloalkyl, C1-6 optionally substituted haloalkyl, C1-6 optionallysubstituted alkoxy, CO₂R⁵, CONR⁶ ₂, NR⁷ ₂, sulfate, sulfonate,optionally substituted aryl, optionally substituted aryloxy, optionallysubstituted hetero aryl, and optionally substituted heterocycle; R^(3a),R^(3b), R^(3c), R^(3d), R^(3e), R^(3f), R^(3g), R^(3h), R^(3i), R^(3j),R^(3k), R^(3l), and R^(3m) at each occurrence are each independentlyselected from the group consisting of hydrogen, C1-6 optionallysubstituted alkyl, C1-6 optionally substituted branched alkyl, C1-6optionally substituted haloalkyl, C1-6 optionally substituted alkoxy,and CO₂R⁸; R^(4a), R^(4b), and R^(4c) at each occurrence are eachindependently selected from the group consisting of hydrogen, C1-6optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C1-6 optionally substituted cyclo alkyl, C1-6 optionallysubstituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶₂, NR⁷ ₂, sulfate, sulfonate, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryl, and optionallysubstituted heterocycle; R^(4a) and R^(4b) at each occurrence on athiophene ring are taken together with the atom to which they are boundto form an optionally substituted ring having from 6 ring atomscontaining 2 oxygen atoms; R⁵ at each occurrence is independentlyselected from the group consisting of hydrogen and optionallysubstituted alkyl; R⁶ at each occurrence is independently selected fromthe group consisting of hydrogen and optionally substituted alkyl; R⁷ ateach occurrence is independently selected from the group consisting ofhydrogen and optionally substituted alkyl; R⁸ at each occurrence isindependently selected from the group consisting of hydrogen andoptionally substituted alkyl; wherein compounds of the followingstructures are excluded:


2. The compound of claim 1, having the formula (II):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein: M is selected from the groupconsisting of manganese, molybdenum, rhenium, iron, ruthenium, osmium,cobalt, rhodium, iridium, nickel, platinum, and copper; X is selectedfrom the group consisting of Cl⁻, PF₆ ⁻, Br⁻, BF₄ ⁻, ClO₄ ⁻, CF₃SO₃ ⁻,and SO₄ ⁻²; n=0, 1, 2, or 3; Lig at each occurrence is independentlyselected from the group consisting of

R¹ is selected from the group consisting of

u is an integer; R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), and R^(2f) ateach occurrence are each independently selected from the groupconsisting of hydrogen, C1-6 optionally substituted alkyl, C1-6optionally substituted branched alkyl, C3-7 optionally substitutedcycloalkyl, C1-6 optionally substituted haloalkyl, C1-6 optionallysubstituted alkoxy, CO₂R⁵, CONR⁶ ₂, NR⁷ ₂, sulfate, sulfonate,optionally substituted aryl, optionally substituted aryloxy, optionallysubstituted heteroaryl, and optionally substituted heterocycle; R^(3a),R^(3b), R^(3c), R^(3d), R^(3e), R^(3f), R^(3g), R^(3h), R^(3i), R^(3j),R^(3k), R^(3l), and R^(3m) at each occurrence are each independentlyselected from the group consisting of hydrogen, C1-6 optionallysubstituted alkyl, C1-6 optionally substituted branched alkyl, C1-6optionally substituted haloalkyl, C1-6 optionally substituted alkoxy,and CO₂R⁸; R^(4a), R^(4b), and R^(4c) at each occurrence are eachindependently selected from the group consisting of hydrogen, C1-6optionally substituted alkyl, C1-6 optionally substituted branchedalkyl, C1-6 optionally substituted cyclo alkyl, C1-6 optionallysubstituted haloalkyl, C1-6 optionally substituted alkoxy, CO₂R⁵, CONR⁶₂, NR⁷ ₂, sulfate, sulfonate, optionally substituted aryl, optionallysubstituted aryloxy, optionally substituted heteroaryl, and optionallysubstituted heterocycle; R^(4a) and R^(4b) at each occurrence on athiophene ring are taken together with the atom to which they are boundto form an optionally substituted ring having from 6 ring atomscontaining 2 oxygen atoms; R⁵ at each occurrence is independentlyselected from the group consisting of hydrogen and optionallysubstituted alkyl; R⁶ at each occurrence is independently selected fromthe group consisting of hydrogen and optionally substituted alkyl; R⁷ ateach occurrence is independently selected from the group consisting ofhydrogen and optionally substituted alkyl; R⁸ at each occurrence isindependently selected from the group consisting of hydrogen andoptionally substituted alkyl; wherein compounds of the followingstructures are excluded:


3. The compound of claim 1, having the formula (III):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof wherein: g=0, 1, 2, 3, 4, or
 5. 4. Thecompound of claim 1, having the formula (IV):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof wherein: h=0, 1, 2, 3, 4, or
 5. 5. Acompound having formula (V):

including hydrates, solvates, pharmaceutically acceptable salts,prodrugs and complexes thereof, wherein: Lig at each occurrence isindependently selected from the group consisting of

t is an integer; q=0, 1, 2, 3, 4 or 5; wherein the following compound isexcluded:


6. A method for treating a disease associated with hyperproliferatingcells, said method comprising administering to a subject an effectiveamount of at least one compound according to claim
 1. 7. The method ofclaim 6, wherein the at least one compound is administered in acomposition further comprising at least one excipient.
 8. A method fortreating a disease associated with hyperproliferating cells, said methodcomprising administering to a subject an effective amount of at leastone compound according to claim
 2. 9. The method of claim 8, wherein theat least one compound is administered in a composition furthercomprising at least one excipient.
 10. A method for treating a diseaseassociated with hyperproliferating cells, said method comprisingadministering to a subject an effective amount of at least one compoundaccording to claim
 3. 11. The method of claim 10, wherein the at leastone compound is administered in a composition further comprising atleast one excipient.
 12. A method for treating a disease associated withhyperproliferating cells, said method comprising administering to asubject an effective amount of at least one compound according to claim4.
 13. The method of claim 12, wherein the at least one compound isadministered in a composition further comprising at least one excipient.14. A method for treating a disease associated with hyperproliferatingcells, said method comprising administering to a subject an effectiveamount of at least one compound according to claim
 5. 15. The method ofclaim 14, wherein the at least one compound is administered in acomposition further comprising at least one excipient.
 16. A method fortreating a disease associated with hyperproliferating cells, said methodcomprising administering to a subject an effective amount of at leastone compound of the structure


17. The method of claim 16, wherein the at least one compound isadministered in a composition further comprising at least one excipient.